96-well plates Search Results


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  • 99
    Millipore 96well plates
    96well Plates, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Bio-Rad 96 well plates
    Confirmation of mutation entry clones by barcoded next-gen sequencing Products of the PCR mutagenesis reaction are cloned into the pDONR223 vector via Gateway BP reaction and transformed into competent DH5α bacterial cells. Transformants are spotted on LB + Spectinomycin plates to isolate single colonies. One plate is shown here as a representation. This scheme illustrates the steps involved in mutation sample preparation and next-generation sequencing confirmation. PCR amplification should be done for each mutant at the single colony level prior to sample preparation (illustrated by 96 well plates M1-M4). If a given gene has multiple mutations to clone ( e.g. , four as shown), these mutation PCR products should be separated into different pools (Steps 35-36). A unique barcode adaptor is added to each pool by ligation (Step 37). All pooled mutation PCR products are then sequence-verified by next-generation sequencing technologies (454-FLX, Illumina Solexa, etc) (Steps 38-41).
    96 Well Plates, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 92/100, based on 221 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Bio-Rad ddpcr 96 well plates
    Confirmation of mutation entry clones by barcoded next-gen sequencing Products of the PCR mutagenesis reaction are cloned into the pDONR223 vector via Gateway BP reaction and transformed into competent DH5α bacterial cells. Transformants are spotted on LB + Spectinomycin plates to isolate single colonies. One plate is shown here as a representation. This scheme illustrates the steps involved in mutation sample preparation and next-generation sequencing confirmation. PCR amplification should be done for each mutant at the single colony level prior to sample preparation (illustrated by 96 well plates M1-M4). If a given gene has multiple mutations to clone ( e.g. , four as shown), these mutation PCR products should be separated into different pools (Steps 35-36). A unique barcode adaptor is added to each pool by ligation (Step 37). All pooled mutation PCR products are then sequence-verified by next-generation sequencing technologies (454-FLX, Illumina Solexa, etc) (Steps 38-41).
    Ddpcr 96 Well Plates, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 99/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    PerkinElmer cytostar 96 well plates
    Confirmation of mutation entry clones by barcoded next-gen sequencing Products of the PCR mutagenesis reaction are cloned into the pDONR223 vector via Gateway BP reaction and transformed into competent DH5α bacterial cells. Transformants are spotted on LB + Spectinomycin plates to isolate single colonies. One plate is shown here as a representation. This scheme illustrates the steps involved in mutation sample preparation and next-generation sequencing confirmation. PCR amplification should be done for each mutant at the single colony level prior to sample preparation (illustrated by 96 well plates M1-M4). If a given gene has multiple mutations to clone ( e.g. , four as shown), these mutation PCR products should be separated into different pools (Steps 35-36). A unique barcode adaptor is added to each pool by ligation (Step 37). All pooled mutation PCR products are then sequence-verified by next-generation sequencing technologies (454-FLX, Illumina Solexa, etc) (Steps 38-41).
    Cytostar 96 Well Plates, supplied by PerkinElmer, used in various techniques. Bioz Stars score: 99/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Avantor 96 well plates
    Confirmation of mutation entry clones by barcoded next-gen sequencing Products of the PCR mutagenesis reaction are cloned into the pDONR223 vector via Gateway BP reaction and transformed into competent DH5α bacterial cells. Transformants are spotted on LB + Spectinomycin plates to isolate single colonies. One plate is shown here as a representation. This scheme illustrates the steps involved in mutation sample preparation and next-generation sequencing confirmation. PCR amplification should be done for each mutant at the single colony level prior to sample preparation (illustrated by 96 well plates M1-M4). If a given gene has multiple mutations to clone ( e.g. , four as shown), these mutation PCR products should be separated into different pools (Steps 35-36). A unique barcode adaptor is added to each pool by ligation (Step 37). All pooled mutation PCR products are then sequence-verified by next-generation sequencing technologies (454-FLX, Illumina Solexa, etc) (Steps 38-41).
    96 Well Plates, supplied by Avantor, used in various techniques. Bioz Stars score: 93/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Axygen 96 well plates
    Confirmation of mutation entry clones by barcoded next-gen sequencing Products of the PCR mutagenesis reaction are cloned into the pDONR223 vector via Gateway BP reaction and transformed into competent DH5α bacterial cells. Transformants are spotted on LB + Spectinomycin plates to isolate single colonies. One plate is shown here as a representation. This scheme illustrates the steps involved in mutation sample preparation and next-generation sequencing confirmation. PCR amplification should be done for each mutant at the single colony level prior to sample preparation (illustrated by 96 well plates M1-M4). If a given gene has multiple mutations to clone ( e.g. , four as shown), these mutation PCR products should be separated into different pools (Steps 35-36). A unique barcode adaptor is added to each pool by ligation (Step 37). All pooled mutation PCR products are then sequence-verified by next-generation sequencing technologies (454-FLX, Illumina Solexa, etc) (Steps 38-41).
    96 Well Plates, supplied by Axygen, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Becton Dickinson 96 well plates
    EF24 shows more potent cytotoxic effect than curcumin on cancer cells. Cells were grown in <t>96-well</t> plates and were treated with EF24 or curcumin as indicated for 48 h. Cell viability was assessed by the sulforhodamine B method and expressed as percentage
    96 Well Plates, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 92/100, based on 410 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    BMG Labtech 96 well plates
    EF24 shows more potent cytotoxic effect than curcumin on cancer cells. Cells were grown in <t>96-well</t> plates and were treated with EF24 or curcumin as indicated for 48 h. Cell viability was assessed by the sulforhodamine B method and expressed as percentage
    96 Well Plates, supplied by BMG Labtech, used in various techniques. Bioz Stars score: 92/100, based on 25 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Boster Bio 96 well plates
    EF24 shows more potent cytotoxic effect than curcumin on cancer cells. Cells were grown in <t>96-well</t> plates and were treated with EF24 or curcumin as indicated for 48 h. Cell viability was assessed by the sulforhodamine B method and expressed as percentage
    96 Well Plates, supplied by Boster Bio, used in various techniques. Bioz Stars score: 93/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Corning Life Sciences 96 well plates
    Antimicrobial screen against E. coli (top) and P. aeruginosa (bottom). ( a ) Bacterial growth kinetics. Growth of these bacteria is expressed as optical density at 600 nm (shown as a.u. (600 nm)) for cultures on <t>96-well</t> plates over 48 h
    96 Well Plates, supplied by Corning Life Sciences, used in various techniques. Bioz Stars score: 92/100, based on 385 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Essen Bioscience 96 well plates
    Antimicrobial screen against E. coli (top) and P. aeruginosa (bottom). ( a ) Bacterial growth kinetics. Growth of these bacteria is expressed as optical density at 600 nm (shown as a.u. (600 nm)) for cultures on <t>96-well</t> plates over 48 h
    96 Well Plates, supplied by Essen Bioscience, used in various techniques. Bioz Stars score: 91/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Greiner Bio 96 well plates
    High-throughput time-resolved microscopy screen for atypical lysis kinetics. (A) Shown is a simplified schematic of the workflow. Cephalexin is added to growing cell cultures in glass-bottom <t>96-well</t> plates and then the plate is kept on the microscope. Images are taken at 4 time-points: 40 min, 85 min, 2 and 4.5 h. Images are then processed to count the number of intact cells at every time-point for each well/strain. Lysed and intact cells are distinguished by thresholding. (B) 96-well plate view of an experiment at T 40 min . Zoomed insets show two wells with different phenotypes. Well marked with ( # ) is representative of most strains, showing intact elongated cells. Well containing the rapidly lysing strain Δ lpoB shows lysed cells at T 40 min . Scale bars correspond to 20 μm. (C) 96-well plate view of an experiment at T 4 . 5h . Zoomed insets show two wells with different phenotypes. Well marked with ( # ) is representative of most strains, showing lysed cells. Well containing the late lysing strain Δ fkpB with intact cells. Scale bars correspond to 100 μm.
    96 Well Plates, supplied by Greiner Bio, used in various techniques. Bioz Stars score: 92/100, based on 156 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Sarstedt 96 well plates
    High-throughput time-resolved microscopy screen for atypical lysis kinetics. (A) Shown is a simplified schematic of the workflow. Cephalexin is added to growing cell cultures in glass-bottom <t>96-well</t> plates and then the plate is kept on the microscope. Images are taken at 4 time-points: 40 min, 85 min, 2 and 4.5 h. Images are then processed to count the number of intact cells at every time-point for each well/strain. Lysed and intact cells are distinguished by thresholding. (B) 96-well plate view of an experiment at T 40 min . Zoomed insets show two wells with different phenotypes. Well marked with ( # ) is representative of most strains, showing intact elongated cells. Well containing the rapidly lysing strain Δ lpoB shows lysed cells at T 40 min . Scale bars correspond to 20 μm. (C) 96-well plate view of an experiment at T 4 . 5h . Zoomed insets show two wells with different phenotypes. Well marked with ( # ) is representative of most strains, showing lysed cells. Well containing the late lysing strain Δ fkpB with intact cells. Scale bars correspond to 100 μm.
    96 Well Plates, supplied by Sarstedt, used in various techniques. Bioz Stars score: 93/100, based on 29 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Iwaki America 96 well plates
    High-throughput time-resolved microscopy screen for atypical lysis kinetics. (A) Shown is a simplified schematic of the workflow. Cephalexin is added to growing cell cultures in glass-bottom <t>96-well</t> plates and then the plate is kept on the microscope. Images are taken at 4 time-points: 40 min, 85 min, 2 and 4.5 h. Images are then processed to count the number of intact cells at every time-point for each well/strain. Lysed and intact cells are distinguished by thresholding. (B) 96-well plate view of an experiment at T 40 min . Zoomed insets show two wells with different phenotypes. Well marked with ( # ) is representative of most strains, showing intact elongated cells. Well containing the rapidly lysing strain Δ lpoB shows lysed cells at T 40 min . Scale bars correspond to 20 μm. (C) 96-well plate view of an experiment at T 4 . 5h . Zoomed insets show two wells with different phenotypes. Well marked with ( # ) is representative of most strains, showing lysed cells. Well containing the late lysing strain Δ fkpB with intact cells. Scale bars correspond to 100 μm.
    96 Well Plates, supplied by Iwaki America, used in various techniques. Bioz Stars score: 93/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Nest Biotechnology 96 well plates
    High-throughput time-resolved microscopy screen for atypical lysis kinetics. (A) Shown is a simplified schematic of the workflow. Cephalexin is added to growing cell cultures in glass-bottom <t>96-well</t> plates and then the plate is kept on the microscope. Images are taken at 4 time-points: 40 min, 85 min, 2 and 4.5 h. Images are then processed to count the number of intact cells at every time-point for each well/strain. Lysed and intact cells are distinguished by thresholding. (B) 96-well plate view of an experiment at T 40 min . Zoomed insets show two wells with different phenotypes. Well marked with ( # ) is representative of most strains, showing intact elongated cells. Well containing the rapidly lysing strain Δ lpoB shows lysed cells at T 40 min . Scale bars correspond to 20 μm. (C) 96-well plate view of an experiment at T 4 . 5h . Zoomed insets show two wells with different phenotypes. Well marked with ( # ) is representative of most strains, showing lysed cells. Well containing the late lysing strain Δ fkpB with intact cells. Scale bars correspond to 100 μm.
    96 Well Plates, supplied by Nest Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 32 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TPP-Techno Plastic 96 well plates
    High-throughput time-resolved microscopy screen for atypical lysis kinetics. (A) Shown is a simplified schematic of the workflow. Cephalexin is added to growing cell cultures in glass-bottom <t>96-well</t> plates and then the plate is kept on the microscope. Images are taken at 4 time-points: 40 min, 85 min, 2 and 4.5 h. Images are then processed to count the number of intact cells at every time-point for each well/strain. Lysed and intact cells are distinguished by thresholding. (B) 96-well plate view of an experiment at T 40 min . Zoomed insets show two wells with different phenotypes. Well marked with ( # ) is representative of most strains, showing intact elongated cells. Well containing the rapidly lysing strain Δ lpoB shows lysed cells at T 40 min . Scale bars correspond to 20 μm. (C) 96-well plate view of an experiment at T 4 . 5h . Zoomed insets show two wells with different phenotypes. Well marked with ( # ) is representative of most strains, showing lysed cells. Well containing the late lysing strain Δ fkpB with intact cells. Scale bars correspond to 100 μm.
    96 Well Plates, supplied by TPP-Techno Plastic, used in various techniques. Bioz Stars score: 93/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    CELLTREAT Scientific 96 well plates
    High-throughput time-resolved microscopy screen for atypical lysis kinetics. (A) Shown is a simplified schematic of the workflow. Cephalexin is added to growing cell cultures in glass-bottom <t>96-well</t> plates and then the plate is kept on the microscope. Images are taken at 4 time-points: 40 min, 85 min, 2 and 4.5 h. Images are then processed to count the number of intact cells at every time-point for each well/strain. Lysed and intact cells are distinguished by thresholding. (B) 96-well plate view of an experiment at T 40 min . Zoomed insets show two wells with different phenotypes. Well marked with ( # ) is representative of most strains, showing intact elongated cells. Well containing the rapidly lysing strain Δ lpoB shows lysed cells at T 40 min . Scale bars correspond to 20 μm. (C) 96-well plate view of an experiment at T 4 . 5h . Zoomed insets show two wells with different phenotypes. Well marked with ( # ) is representative of most strains, showing lysed cells. Well containing the late lysing strain Δ fkpB with intact cells. Scale bars correspond to 100 μm.
    96 Well Plates, supplied by CELLTREAT Scientific, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Dynatech Laboratories 96 well plates
    High-throughput time-resolved microscopy screen for atypical lysis kinetics. (A) Shown is a simplified schematic of the workflow. Cephalexin is added to growing cell cultures in glass-bottom <t>96-well</t> plates and then the plate is kept on the microscope. Images are taken at 4 time-points: 40 min, 85 min, 2 and 4.5 h. Images are then processed to count the number of intact cells at every time-point for each well/strain. Lysed and intact cells are distinguished by thresholding. (B) 96-well plate view of an experiment at T 40 min . Zoomed insets show two wells with different phenotypes. Well marked with ( # ) is representative of most strains, showing intact elongated cells. Well containing the rapidly lysing strain Δ lpoB shows lysed cells at T 40 min . Scale bars correspond to 20 μm. (C) 96-well plate view of an experiment at T 4 . 5h . Zoomed insets show two wells with different phenotypes. Well marked with ( # ) is representative of most strains, showing lysed cells. Well containing the late lysing strain Δ fkpB with intact cells. Scale bars correspond to 100 μm.
    96 Well Plates, supplied by Dynatech Laboratories, used in various techniques. Bioz Stars score: 92/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    MJ Research 96 well plates
    High-throughput time-resolved microscopy screen for atypical lysis kinetics. (A) Shown is a simplified schematic of the workflow. Cephalexin is added to growing cell cultures in glass-bottom <t>96-well</t> plates and then the plate is kept on the microscope. Images are taken at 4 time-points: 40 min, 85 min, 2 and 4.5 h. Images are then processed to count the number of intact cells at every time-point for each well/strain. Lysed and intact cells are distinguished by thresholding. (B) 96-well plate view of an experiment at T 40 min . Zoomed insets show two wells with different phenotypes. Well marked with ( # ) is representative of most strains, showing intact elongated cells. Well containing the rapidly lysing strain Δ lpoB shows lysed cells at T 40 min . Scale bars correspond to 20 μm. (C) 96-well plate view of an experiment at T 4 . 5h . Zoomed insets show two wells with different phenotypes. Well marked with ( # ) is representative of most strains, showing lysed cells. Well containing the late lysing strain Δ fkpB with intact cells. Scale bars correspond to 100 μm.
    96 Well Plates, supplied by MJ Research, used in various techniques. Bioz Stars score: 92/100, based on 41 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    Promega 96 well plates
    Subcutaneous and visceral adipose-derived mesenchymal stem cells (ASCs) display comparable cell surface marker profiles, cell cycle distribution and cell proliferation. ( A ) Immunofluorescence staining of mesenchymal stem cell surface markers CD90 (green) and CD73 (red), and DNA (DAPI, blue) in subcutaneous ASCs (ASCsub) and visceral ASCs (ASCvis). Scale: 20 μm. ( B ) Flow cytometric analyses of positive cell surface markers CD90, CD73, CD146, and CD105, and negative markers CD14, CD31, CD106, and CD34 for mesenchymal stem cells (MSCs). Values represent the percentages of ASCs expressing the indicated protein. The results from eight independent experiments (donors) are presented as mean ± standard error of the mean (SEM). ( C , D ) Cell cycle distribution was analyzed using a FACSCalibur TM . Profile examples were shown ( C ). Cell cycle phases of ASCs were presented in percentage and the results were derived from four independent experiments ( D ). ( E , F ) ASCs were stained for pHH3 (S10) (green), α-tubulin (yellow), pericentrin (red) and DNA (blue), and representatives are shown ( E ). Scale: 10 μm. pHH3 positive cells were quantified in ASCsub and ASCvis ( F ). The results are from three independent experiments with ASCs from three different donors and presented as median ± min/max whiskers in box plots. n.s. > 0.05. ( G ) Cellular extracts from ASCs were prepared for Western blot analyses with indicated antibodies. β-actin served as loading control. ( H ) ASCs were seeded in <t>96-well</t> plates for 0, 24, 48, 72, and 96 h. Cell viability was measured via CellTiter-Blue ® assay. The results are presented as mean ± SEM and statistically analyzed, showing no significant difference (n.s.).
    96 Well Plates, supplied by Promega, used in various techniques. Bioz Stars score: 96/100, based on 173 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    SPL Life Sciences 96 well plates
    Cultivability of the four Bacillus subtilis sub-populations after high pressure treatment. Spores were treated at 150 MPa and 37°C for 10 min in 50 mM ACES buffer at pH 7, stained with SYTO16 and PI. Four sub-populations were observed with the following presumptive assignment: R1: superdormant spores, R2: germinated spores, R3: previously referred to as “unknown”, R4: membrane-compromised spores. Each sub-population was single cell sorted into <t>96-well</t> plates containing tryptic soy agar through FACS and incubated at 37°C for 48 h. Error bars present standard deviations of three independent treatments ( n = 3).
    96 Well Plates, supplied by SPL Life Sciences, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Fisher Scientific 96 well plates
    Cultivability of the four Bacillus subtilis sub-populations after high pressure treatment. Spores were treated at 150 MPa and 37°C for 10 min in 50 mM ACES buffer at pH 7, stained with SYTO16 and PI. Four sub-populations were observed with the following presumptive assignment: R1: superdormant spores, R2: germinated spores, R3: previously referred to as “unknown”, R4: membrane-compromised spores. Each sub-population was single cell sorted into <t>96-well</t> plates containing tryptic soy agar through FACS and incubated at 37°C for 48 h. Error bars present standard deviations of three independent treatments ( n = 3).
    96 Well Plates, supplied by Fisher Scientific, used in various techniques. Bioz Stars score: 92/100, based on 40 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Confirmation of mutation entry clones by barcoded next-gen sequencing Products of the PCR mutagenesis reaction are cloned into the pDONR223 vector via Gateway BP reaction and transformed into competent DH5α bacterial cells. Transformants are spotted on LB + Spectinomycin plates to isolate single colonies. One plate is shown here as a representation. This scheme illustrates the steps involved in mutation sample preparation and next-generation sequencing confirmation. PCR amplification should be done for each mutant at the single colony level prior to sample preparation (illustrated by 96 well plates M1-M4). If a given gene has multiple mutations to clone ( e.g. , four as shown), these mutation PCR products should be separated into different pools (Steps 35-36). A unique barcode adaptor is added to each pool by ligation (Step 37). All pooled mutation PCR products are then sequence-verified by next-generation sequencing technologies (454-FLX, Illumina Solexa, etc) (Steps 38-41).

    Journal: Nature protocols

    Article Title: Base-resolution stratification of cancer mutations using functional variomics

    doi: 10.1038/nprot.2017.086

    Figure Lengend Snippet: Confirmation of mutation entry clones by barcoded next-gen sequencing Products of the PCR mutagenesis reaction are cloned into the pDONR223 vector via Gateway BP reaction and transformed into competent DH5α bacterial cells. Transformants are spotted on LB + Spectinomycin plates to isolate single colonies. One plate is shown here as a representation. This scheme illustrates the steps involved in mutation sample preparation and next-generation sequencing confirmation. PCR amplification should be done for each mutant at the single colony level prior to sample preparation (illustrated by 96 well plates M1-M4). If a given gene has multiple mutations to clone ( e.g. , four as shown), these mutation PCR products should be separated into different pools (Steps 35-36). A unique barcode adaptor is added to each pool by ligation (Step 37). All pooled mutation PCR products are then sequence-verified by next-generation sequencing technologies (454-FLX, Illumina Solexa, etc) (Steps 38-41).

    Article Snippet: Round-bottom 96-well plates (Corning, cat. no. 3788) 50 ml sterile reagent troughs (Corning, cat. no. 4871) Thin-walled 96-well PCR microtiter plates (Bio-Rad, cat. no. HSP-9601) 15 cm sterile petri dishes (Fisher Scientific, cat. no. 08-757-14) Aluminum foil microplate seal (Bio-Rad, cat. no. MSF-1001) 50 ml polypropylene tubes (BD Falcon, cat. no. 352070) Tissue culture 96 wells flat bottom plate (BD Falcon, cat. no. 353075) PIPETMAN P10, 1 to 10 µl (Gibson, cat. no. F144802) PIPETMAN P200, 50 to 200 µl (Gibson, cat. no. F123601) Corning™ 4084 P10 multichannel pipettes 1 - 10 µl (Thermofisher scientific, cat. no. 07-764-710) Corning™ 4085 P50 multichannel pipettes 5 - 50 µl (Thermofisher scientific, cat. no. 07-764-711) Corning™ 4085 P200 multichannel pipettes 20 - 200 µl (Thermofisher scientific, cat. no. 07-764-712) Corning® Stripettor™ Ultra Pipet Controller (Thermofisher scientific, cat. no. 4099) Thermocycler capable of handling 96-well plates (Bio-Rad, cat. no. T100™) Temperature controlled incubator (30°C) (Thermofisher scientific, cat. no. 15-103-0515) Temperature controlled water bath (42°C) (Thermofisher scientific, cat. no. FSGPD10) 1% E-Gel® 96 Agarose Gels (Thermofisher scientific, cat. no. G700801) E-gel Mother E-Base capable of running 96 samples (Thermofisher scientific, cat. no. EBM03) Temperature controlled shaker for 250 ml flasks and 50 ml tubes (MaxQ™ 6000 Shaker, Thermofisher scientific, cat. no. SHKE6000) Benchtop Centrifuge suitable for microplates and 50 ml tubes (Sorvall™ Legend™ XT/XF centrifuge, Thermofisher scientific, cat. no. 75-004-505) Plate shaker (Mixmate, Eppendorf, cat. no. 022674200) GENESYS™ 10S UV-Vis Spectrophotometer (Thermofisher scientific, cat. no. 840-208100) Vortexer (Thermofisher scientific, cat. no. 88880017TS) Centro XS3 Luminescence Microplate Reader (Berthold Technologies, cat. no. 46970) 1300 Series A2 Class II, Type A2 Bio Safety Cabinets (Thermofisher scientific, cat. no. 13-261-221) Optional: liquid handling robot with 96-well multichannel head and individual well cherry-picking capability (Tecan, cat. no. Fluent 780) Optional: BioRobot Universal System Robot with 96-well DNA miniprep capability (Qiagen, cat. no. 9001094)

    Techniques: Mutagenesis, Clone Assay, Sequencing, Polymerase Chain Reaction, Plasmid Preparation, Transformation Assay, Sample Prep, Next-Generation Sequencing, Amplification, Ligation

    EF24 shows more potent cytotoxic effect than curcumin on cancer cells. Cells were grown in 96-well plates and were treated with EF24 or curcumin as indicated for 48 h. Cell viability was assessed by the sulforhodamine B method and expressed as percentage

    Journal:

    Article Title: Inhibition of IκB Kinase-Nuclear Factor-κB Signaling Pathway by 3,5-Bis(2-flurobenzylidene)piperidin-4-one (EF24), a Novel Monoketone Analog of Curcumin *

    doi: 10.1124/mol.108.046201

    Figure Lengend Snippet: EF24 shows more potent cytotoxic effect than curcumin on cancer cells. Cells were grown in 96-well plates and were treated with EF24 or curcumin as indicated for 48 h. Cell viability was assessed by the sulforhodamine B method and expressed as percentage

    Article Snippet: A549 cells were plated in 96-well plates (BD Biosciences Discovery Labware, Bedford, MA) at 10,000 cells/90 μ l/well and grown for 20 h. Test compounds were added to each well and incubated at 37°C.

    Techniques:

    EF24 impairs TNF- α induced NF- κ B nuclear translocation. A549 cells were grown in 96-well plates and treated with TNF- α or control (DMSO) for 30 min before sample processing for the detection of NF- κ B as described under

    Journal:

    Article Title: Inhibition of IκB Kinase-Nuclear Factor-κB Signaling Pathway by 3,5-Bis(2-flurobenzylidene)piperidin-4-one (EF24), a Novel Monoketone Analog of Curcumin *

    doi: 10.1124/mol.108.046201

    Figure Lengend Snippet: EF24 impairs TNF- α induced NF- κ B nuclear translocation. A549 cells were grown in 96-well plates and treated with TNF- α or control (DMSO) for 30 min before sample processing for the detection of NF- κ B as described under

    Article Snippet: A549 cells were plated in 96-well plates (BD Biosciences Discovery Labware, Bedford, MA) at 10,000 cells/90 μ l/well and grown for 20 h. Test compounds were added to each well and incubated at 37°C.

    Techniques: Translocation Assay

    Antimicrobial screen against E. coli (top) and P. aeruginosa (bottom). ( a ) Bacterial growth kinetics. Growth of these bacteria is expressed as optical density at 600 nm (shown as a.u. (600 nm)) for cultures on 96-well plates over 48 h

    Journal:

    Article Title: Small molecules with antimicrobial activity against E. coli and P. aeruginosa identified by high-throughput screening

    doi: 10.1038/sj.bjp.0706873

    Figure Lengend Snippet: Antimicrobial screen against E. coli (top) and P. aeruginosa (bottom). ( a ) Bacterial growth kinetics. Growth of these bacteria is expressed as optical density at 600 nm (shown as a.u. (600 nm)) for cultures on 96-well plates over 48 h

    Article Snippet: Compounds were in 96-well plates (Corning-Costar) as 10 m M solutions in dimethylsulphoxide (DMSO).

    Techniques:

    High-throughput time-resolved microscopy screen for atypical lysis kinetics. (A) Shown is a simplified schematic of the workflow. Cephalexin is added to growing cell cultures in glass-bottom 96-well plates and then the plate is kept on the microscope. Images are taken at 4 time-points: 40 min, 85 min, 2 and 4.5 h. Images are then processed to count the number of intact cells at every time-point for each well/strain. Lysed and intact cells are distinguished by thresholding. (B) 96-well plate view of an experiment at T 40 min . Zoomed insets show two wells with different phenotypes. Well marked with ( # ) is representative of most strains, showing intact elongated cells. Well containing the rapidly lysing strain Δ lpoB shows lysed cells at T 40 min . Scale bars correspond to 20 μm. (C) 96-well plate view of an experiment at T 4 . 5h . Zoomed insets show two wells with different phenotypes. Well marked with ( # ) is representative of most strains, showing lysed cells. Well containing the late lysing strain Δ fkpB with intact cells. Scale bars correspond to 100 μm.

    Journal: Frontiers in Microbiology

    Article Title: Image-Based Dynamic Phenotyping Reveals Genetic Determinants of Filamentation-Mediated β-Lactam Tolerance

    doi: 10.3389/fmicb.2020.00374

    Figure Lengend Snippet: High-throughput time-resolved microscopy screen for atypical lysis kinetics. (A) Shown is a simplified schematic of the workflow. Cephalexin is added to growing cell cultures in glass-bottom 96-well plates and then the plate is kept on the microscope. Images are taken at 4 time-points: 40 min, 85 min, 2 and 4.5 h. Images are then processed to count the number of intact cells at every time-point for each well/strain. Lysed and intact cells are distinguished by thresholding. (B) 96-well plate view of an experiment at T 40 min . Zoomed insets show two wells with different phenotypes. Well marked with ( # ) is representative of most strains, showing intact elongated cells. Well containing the rapidly lysing strain Δ lpoB shows lysed cells at T 40 min . Scale bars correspond to 20 μm. (C) 96-well plate view of an experiment at T 4 . 5h . Zoomed insets show two wells with different phenotypes. Well marked with ( # ) is representative of most strains, showing lysed cells. Well containing the late lysing strain Δ fkpB with intact cells. Scale bars correspond to 100 μm.

    Article Snippet: Briefly, strains were grown overnight in 96-well plates (Greiner, 96 Well Polystyrene Microplate, clear) containing LB with 30 μg/ml Kanamycin (MP Biomedicals), and then diluted 5000 times in a 96-well plate with glass bottom (Brooks Automation Ltd., Ref. MGB096-1-2-LG-L) containing fresh media.

    Techniques: High Throughput Screening Assay, Microscopy, Lysis

    Measuring β-lactam induced lysis kinetics by phase contrast microscopy. (A) Time-lapse analysis of wild-type cells on agar pads containing cephalexin 100 μg/ml. Cells immediately stop dividing after seeded on the cephalexin containing agar but continue to elongate. After growing to variable lengths, the cells develop a bulge (shown by red arrow heads) and lyse. Scale bar corresponds to 25 μm. (B) Graph shows wild-type E. coli cell counts at different time intervals after treatment with cephalexin 100 μg/ml at t = 0 in glass-bottom imaging microplates. Cell counts were determined simultaneously by image analysis (left axis in red) and by CFU counting on agar plates (right axis in blue). (C) Wild-type E. coli was treated with cephalexin 200 μg/ml in glass-bottom 96-well plates at different cell densities and cell counts were determined based on image analysis. All data points correspond to 3 replicates in (B,C) .

    Journal: Frontiers in Microbiology

    Article Title: Image-Based Dynamic Phenotyping Reveals Genetic Determinants of Filamentation-Mediated β-Lactam Tolerance

    doi: 10.3389/fmicb.2020.00374

    Figure Lengend Snippet: Measuring β-lactam induced lysis kinetics by phase contrast microscopy. (A) Time-lapse analysis of wild-type cells on agar pads containing cephalexin 100 μg/ml. Cells immediately stop dividing after seeded on the cephalexin containing agar but continue to elongate. After growing to variable lengths, the cells develop a bulge (shown by red arrow heads) and lyse. Scale bar corresponds to 25 μm. (B) Graph shows wild-type E. coli cell counts at different time intervals after treatment with cephalexin 100 μg/ml at t = 0 in glass-bottom imaging microplates. Cell counts were determined simultaneously by image analysis (left axis in red) and by CFU counting on agar plates (right axis in blue). (C) Wild-type E. coli was treated with cephalexin 200 μg/ml in glass-bottom 96-well plates at different cell densities and cell counts were determined based on image analysis. All data points correspond to 3 replicates in (B,C) .

    Article Snippet: Briefly, strains were grown overnight in 96-well plates (Greiner, 96 Well Polystyrene Microplate, clear) containing LB with 30 μg/ml Kanamycin (MP Biomedicals), and then diluted 5000 times in a 96-well plate with glass bottom (Brooks Automation Ltd., Ref. MGB096-1-2-LG-L) containing fresh media.

    Techniques: Lysis, Microscopy, Imaging

    Subcutaneous and visceral adipose-derived mesenchymal stem cells (ASCs) display comparable cell surface marker profiles, cell cycle distribution and cell proliferation. ( A ) Immunofluorescence staining of mesenchymal stem cell surface markers CD90 (green) and CD73 (red), and DNA (DAPI, blue) in subcutaneous ASCs (ASCsub) and visceral ASCs (ASCvis). Scale: 20 μm. ( B ) Flow cytometric analyses of positive cell surface markers CD90, CD73, CD146, and CD105, and negative markers CD14, CD31, CD106, and CD34 for mesenchymal stem cells (MSCs). Values represent the percentages of ASCs expressing the indicated protein. The results from eight independent experiments (donors) are presented as mean ± standard error of the mean (SEM). ( C , D ) Cell cycle distribution was analyzed using a FACSCalibur TM . Profile examples were shown ( C ). Cell cycle phases of ASCs were presented in percentage and the results were derived from four independent experiments ( D ). ( E , F ) ASCs were stained for pHH3 (S10) (green), α-tubulin (yellow), pericentrin (red) and DNA (blue), and representatives are shown ( E ). Scale: 10 μm. pHH3 positive cells were quantified in ASCsub and ASCvis ( F ). The results are from three independent experiments with ASCs from three different donors and presented as median ± min/max whiskers in box plots. n.s. > 0.05. ( G ) Cellular extracts from ASCs were prepared for Western blot analyses with indicated antibodies. β-actin served as loading control. ( H ) ASCs were seeded in 96-well plates for 0, 24, 48, 72, and 96 h. Cell viability was measured via CellTiter-Blue ® assay. The results are presented as mean ± SEM and statistically analyzed, showing no significant difference (n.s.).

    Journal: Cells

    Article Title: Subcutaneous and Visceral Adipose-Derived Mesenchymal Stem Cells: Commonality and Diversity

    doi: 10.3390/cells8101288

    Figure Lengend Snippet: Subcutaneous and visceral adipose-derived mesenchymal stem cells (ASCs) display comparable cell surface marker profiles, cell cycle distribution and cell proliferation. ( A ) Immunofluorescence staining of mesenchymal stem cell surface markers CD90 (green) and CD73 (red), and DNA (DAPI, blue) in subcutaneous ASCs (ASCsub) and visceral ASCs (ASCvis). Scale: 20 μm. ( B ) Flow cytometric analyses of positive cell surface markers CD90, CD73, CD146, and CD105, and negative markers CD14, CD31, CD106, and CD34 for mesenchymal stem cells (MSCs). Values represent the percentages of ASCs expressing the indicated protein. The results from eight independent experiments (donors) are presented as mean ± standard error of the mean (SEM). ( C , D ) Cell cycle distribution was analyzed using a FACSCalibur TM . Profile examples were shown ( C ). Cell cycle phases of ASCs were presented in percentage and the results were derived from four independent experiments ( D ). ( E , F ) ASCs were stained for pHH3 (S10) (green), α-tubulin (yellow), pericentrin (red) and DNA (blue), and representatives are shown ( E ). Scale: 10 μm. pHH3 positive cells were quantified in ASCsub and ASCvis ( F ). The results are from three independent experiments with ASCs from three different donors and presented as median ± min/max whiskers in box plots. n.s. > 0.05. ( G ) Cellular extracts from ASCs were prepared for Western blot analyses with indicated antibodies. β-actin served as loading control. ( H ) ASCs were seeded in 96-well plates for 0, 24, 48, 72, and 96 h. Cell viability was measured via CellTiter-Blue ® assay. The results are presented as mean ± SEM and statistically analyzed, showing no significant difference (n.s.).

    Article Snippet: Cell proliferation assays were carried out by using Cell Titer-Blue® Cell Viability Assay (BD Biosciences, Heidelberg, Germany) on treated cells in 96-well plates (Promega, Mannheim, Germany).

    Techniques: Derivative Assay, Marker, Immunofluorescence, Staining, Flow Cytometry, Expressing, Western Blot, CtB Assay

    Cultivability of the four Bacillus subtilis sub-populations after high pressure treatment. Spores were treated at 150 MPa and 37°C for 10 min in 50 mM ACES buffer at pH 7, stained with SYTO16 and PI. Four sub-populations were observed with the following presumptive assignment: R1: superdormant spores, R2: germinated spores, R3: previously referred to as “unknown”, R4: membrane-compromised spores. Each sub-population was single cell sorted into 96-well plates containing tryptic soy agar through FACS and incubated at 37°C for 48 h. Error bars present standard deviations of three independent treatments ( n = 3).

    Journal: Frontiers in Microbiology

    Article Title: Flow Cytometry Combined With Single Cell Sorting to Study Heterogeneous Germination of Bacillus Spores Under High Pressure

    doi: 10.3389/fmicb.2019.03118

    Figure Lengend Snippet: Cultivability of the four Bacillus subtilis sub-populations after high pressure treatment. Spores were treated at 150 MPa and 37°C for 10 min in 50 mM ACES buffer at pH 7, stained with SYTO16 and PI. Four sub-populations were observed with the following presumptive assignment: R1: superdormant spores, R2: germinated spores, R3: previously referred to as “unknown”, R4: membrane-compromised spores. Each sub-population was single cell sorted into 96-well plates containing tryptic soy agar through FACS and incubated at 37°C for 48 h. Error bars present standard deviations of three independent treatments ( n = 3).

    Article Snippet: Treated spores were sorted into 96-well plates (SPL Life Sciences, Pocheon, South Korea) filled with TSA.

    Techniques: Staining, FACS, Incubation

    Cultivability of stained and unstained high pressure treated Bacillus subtilis spores. Spores were high pressure treated at 150 MPa and 37°C for 10 min in 50 mM ACES buffer at pH 7, left unstained or stained with SYTO16 and PI, and single cell sorted into a 96-well plate containing tryptic soy agar through FACS. Error bars present standard deviations ( n = 3).

    Journal: Frontiers in Microbiology

    Article Title: Flow Cytometry Combined With Single Cell Sorting to Study Heterogeneous Germination of Bacillus Spores Under High Pressure

    doi: 10.3389/fmicb.2019.03118

    Figure Lengend Snippet: Cultivability of stained and unstained high pressure treated Bacillus subtilis spores. Spores were high pressure treated at 150 MPa and 37°C for 10 min in 50 mM ACES buffer at pH 7, left unstained or stained with SYTO16 and PI, and single cell sorted into a 96-well plate containing tryptic soy agar through FACS. Error bars present standard deviations ( n = 3).

    Article Snippet: Treated spores were sorted into 96-well plates (SPL Life Sciences, Pocheon, South Korea) filled with TSA.

    Techniques: Staining, FACS

    Heterogeneous colony growth of FACS single cell sorted high pressure treated Bacillus subtilis spores. Spores were high pressure treated at 150 MPa and 37°C for 10 min in 50 mM ACES buffer at pH 7, and single cells were sorted into a 96-well plate containing tryptic soy agar using FACS. Some wells displayed no growth, e.g., well A4 (non-cultivable), while other wells showed growth, e.g., well A1 (cultivable). Among the cultivable cells, large differences in colony size were observed, e.g., well H7 (large), H8 (middle) and H9 (small).

    Journal: Frontiers in Microbiology

    Article Title: Flow Cytometry Combined With Single Cell Sorting to Study Heterogeneous Germination of Bacillus Spores Under High Pressure

    doi: 10.3389/fmicb.2019.03118

    Figure Lengend Snippet: Heterogeneous colony growth of FACS single cell sorted high pressure treated Bacillus subtilis spores. Spores were high pressure treated at 150 MPa and 37°C for 10 min in 50 mM ACES buffer at pH 7, and single cells were sorted into a 96-well plate containing tryptic soy agar using FACS. Some wells displayed no growth, e.g., well A4 (non-cultivable), while other wells showed growth, e.g., well A1 (cultivable). Among the cultivable cells, large differences in colony size were observed, e.g., well H7 (large), H8 (middle) and H9 (small).

    Article Snippet: Treated spores were sorted into 96-well plates (SPL Life Sciences, Pocheon, South Korea) filled with TSA.

    Techniques: FACS

    Distribution of cultivability of high pressure treated Bacillus subtilis spores. Spores were high pressure treated at 150 MPa and 37°C for 10 min in 50 mM ACES buffer at pH 7, stained with SYTO16 and PI, and single cell sorted into 96-well plates filled with tryptic soy agar through FACS. After 48 h of incubation, colony formation in three 96-well plates, in total 288 wells, was recorded as “cultivable” or “non-cultivable” depending on the presence or absence of a colony in each well. SYTO16 and PI signals recorded during the sorting using the index sorting function are plotted to visualize the distribution of cultivability. Blue dots and red triangles represent cultivable and non-cultivable events, respectively. Dashed lines indicate approximate gate locations of the four presumptive sub-populations.

    Journal: Frontiers in Microbiology

    Article Title: Flow Cytometry Combined With Single Cell Sorting to Study Heterogeneous Germination of Bacillus Spores Under High Pressure

    doi: 10.3389/fmicb.2019.03118

    Figure Lengend Snippet: Distribution of cultivability of high pressure treated Bacillus subtilis spores. Spores were high pressure treated at 150 MPa and 37°C for 10 min in 50 mM ACES buffer at pH 7, stained with SYTO16 and PI, and single cell sorted into 96-well plates filled with tryptic soy agar through FACS. After 48 h of incubation, colony formation in three 96-well plates, in total 288 wells, was recorded as “cultivable” or “non-cultivable” depending on the presence or absence of a colony in each well. SYTO16 and PI signals recorded during the sorting using the index sorting function are plotted to visualize the distribution of cultivability. Blue dots and red triangles represent cultivable and non-cultivable events, respectively. Dashed lines indicate approximate gate locations of the four presumptive sub-populations.

    Article Snippet: Treated spores were sorted into 96-well plates (SPL Life Sciences, Pocheon, South Korea) filled with TSA.

    Techniques: Staining, FACS, Incubation