Journal: Scientific Reports

Article Title: Thioflavin-T: application as a neuronal body and nucleolar stain and the blue light photo enhancement effect

doi: 10.1038/s41598-024-74359-8

Figure Lengend Snippet: Experimental reagents.

Article Snippet: Thioflavin T (THT) , (MedChemExpress) , HY-D0218.

Techniques: Microscopy, Cell Culture

Journal: Scientific Reports

Article Title: Thioflavin-T: application as a neuronal body and nucleolar stain and the blue light photo enhancement effect

doi: 10.1038/s41598-024-74359-8

Figure Lengend Snippet: Representative staining of a mouse brain using THT and NTDR following protocol 1 at 10 × magnification. ( A , B ) THT intensity with 488 nm laser excitation in green pseudo color image ( A ) and heat map ( B ). ( C , D ) NTDR image with a 633 nm laser ( C ) and merge with THT ( D ). Scale bar: 1000 µm.

Article Snippet: Thioflavin T (THT) , (MedChemExpress) , HY-D0218.

Techniques: Staining

Journal: Scientific Reports

Article Title: Thioflavin-T: application as a neuronal body and nucleolar stain and the blue light photo enhancement effect

doi: 10.1038/s41598-024-74359-8

Figure Lengend Snippet: Co-localization analyses of brain regions stained with THT and NTDR, ( A ) Representative images of various brain regions in the brain section stained with NTDR, THT and Hoechst 33342. Scale bar: 20 µm. ( B ) Representative image of the CA3 region of the hippocampus for the intensity line profiles. ( C ) Comparison of line profiles between THT, NTDR and Hoechst 33342. Red: NeuroTrace DR (NTDR), Green: THT, Blue: Hoechst33342. Black brackets indicate the co-localization of NTDR and THT. ( D ) Pearson’s correlation analysis between THT and NTDR in various brain regions n = 5/6, ( E ) Spearman’s Rank Coefficient analysis for each of the brain regions (n = 6 all regions except Cerebellum & hippocampus n = 5, Average of all PCC and SRCC results respectively, an average score of > 0.7 was considered as evidence of co-localization for the PCC and SRCC, ( F ) Tabulated average SRCC and PCC corresponding to ( E ). A one sample T-test was conducted for each brain region in D,E.

Article Snippet: Thioflavin T (THT) , (MedChemExpress) , HY-D0218.

Techniques: Staining, Comparison

Journal: Scientific Reports

Article Title: Thioflavin-T: application as a neuronal body and nucleolar stain and the blue light photo enhancement effect

doi: 10.1038/s41598-024-74359-8

Figure Lengend Snippet: THT as a nucleolar stain in fixed tissue and live cells, ( A ) Representative images of the hippocampus. Blue: Hoechst33342, Red: NTDR, Green: THT. ( B - E ) Cropped image at white square area shown in ( A ). ( B ) Hoechst 33342 with arrows indicating chromatin structures. ( C ) Cropped merge image. ( D ) NTDR with arrow indicating faint nucleolus. ( E ) THT at 488 nm excitation with arrow indicating nucleolus. ( F ) Representative images of the cortex layers III-IV. Blue: Hoechst33342, Red: NTDR, Green: THT. ( G – J ) Cropped image at white square area shown in ( F ). ( G ) Hoechst 33342 with arrows indicating chromatin structures. ( H ) Cropped merge image. ( I ) NTDR with arrow indicating nucleolus. ( J ) THT at 488 nm excitation with arrow indicating nucleolus. ( K , L ) Representative spectral intensity profile of an example nucleus in the white region of ( H ), ( M , N ) Representative image of live BV2 microglia ( M ) and C8-D1A astrocytes ( N ) staining with Hoechst 33342 and THT.

Article Snippet: Thioflavin T (THT) , (MedChemExpress) , HY-D0218.

Techniques: Staining

Journal: Scientific Reports

Article Title: Thioflavin-T: application as a neuronal body and nucleolar stain and the blue light photo enhancement effect

doi: 10.1038/s41598-024-74359-8

Figure Lengend Snippet: THT is compatible with antibody based fluorescent techniques. ( A , B ) Representative images of areas stained with THT, IBA1 (microglia/macrophage marker) and Hoechst 33342 at CA3 region ( A ) and Cortex ( B ). ( C ) Cortex imaged once with a 405 nm laser and then imaged again with THT, NTDR and Hoechst 33342 with THT excited by both 488 and 458 lasers. ( D ) Cortex stained with THT, NTDR and Hoechst 33342 demonstrating the spectral intensity overlap between THT excited with the 458 nm laser and NTDR, ( E ) Hippocampus stained with IBA1, GFAP, THT and Hoechst 33342, 40 × magnification, white arrows indicate IBA1 + microglia that partially stain positive for THT, red arrows indicate GFAP + Astrocytes that microglia that partially stain positive for THT, ( F ) SRCC and PCC analyses for THT vs IBA1 or GFAP with corresponding mean values, pooled data from 2 experiments each with n = 3, Excitation laser wavelengths appear beside the stain used. A one sample T-Test was conducted for F.

Article Snippet: Thioflavin T (THT) , (MedChemExpress) , HY-D0218.

Techniques: Staining, Marker

Journal: Scientific Reports

Article Title: Thioflavin-T: application as a neuronal body and nucleolar stain and the blue light photo enhancement effect

doi: 10.1038/s41598-024-74359-8

Figure Lengend Snippet: THT photo-enhancement by blue-light exposure. ( A , B ) Representative 2.5D images of THT-stained cortex imaged using the 488 nm laser after the exposure of right half side to a 405 nm laser scanning (770 µW) for 10 s ( A ) and 30 s, 40 × magnification ( B ). ( C ) Hippocampus stained with THT, NTDR and Hoechst 33,=342 (each 10 × magnification) imaged immediately after the 405 nm-laser-induced photo-enhancement at 3 regions on the hippocampus using 40 × magnification. ( D , E ) Sample stained only with THT (D) or Hoechst 33342 ( E ) imaged with the 488 nm laser without/with the exposure of the 405 nm laser for 10 s. ( F ) Hippocampus previously exposed to 405 nm laser by 40 × magnification 11 days prior to imaging with a 488 nm excitation laser and 10 × magnification. ( G ) Sample previously exposed to 405 nm laser by 40 × magnification 10 days prior to imaging with the 488 nm excitation laser and 40 × magnification.

Article Snippet: Thioflavin T (THT) , (MedChemExpress) , HY-D0218.

Techniques: Staining, Imaging

Journal: Scientific Reports

Article Title: Thioflavin-T: application as a neuronal body and nucleolar stain and the blue light photo enhancement effect

doi: 10.1038/s41598-024-74359-8

Figure Lengend Snippet: THT blue light photo enhancement and photo bleaching. ( A ) Change in emission intensity of cortex sample calculated from the images shown in ( B ), n = 3 pooled from 2 experiments. ( B ) Representative heat maps acquired by 458 nm/488 nm excitation laser before and after the 405 nm laser illumination (2%, 770 µW) for 10s by 40 × magnification lens. ( C ) Representative heat maps acquired by 458 nm/488 nm excitation laser before and after the 405 nm laser illumination (0.2%, 77 µW) for 10 s by 40 × magnification. ( D ) Representative heat maps acquired by 458 nm/488 nm excitation laser before and after the 365 nm halogen lamp (25%) for 60 s by 49wf blue filter (Carl Zeiss) and 40 × magnification. Arrows represent examples where loss of signal is evident after blue light exposure in the using 458 nm. A 2 sample T-test was conducted for A.

Article Snippet: Thioflavin T (THT) , (MedChemExpress) , HY-D0218.

Techniques:

Journal: Scientific Reports

Article Title: Thioflavin-T: application as a neuronal body and nucleolar stain and the blue light photo enhancement effect

doi: 10.1038/s41598-024-74359-8

Figure Lengend Snippet: Establishment of THT staining protocols, ( A ) Illustration of THT staining protocols A and B with accompanying notes. ( B ) Representative image of a female 4 month-old C57B/6 mouse brain section viewed on the sagittal plane stained using protocol A imaged using the 458 nm excitation laser and 10 × magnification. ( C ) Representative image of a female 14 month-old APP NL-GLF brain section viewed on the sagittal plane stained using protocol B imaged using the 458 nm excitation laser and 10 × magnification lens with digital zoom to highlight the cortex and hippocampus. ( D ) Representative image of a THT-stained plaque in the molecular layer surround the dentate gyrus taken at 40 × magnification (left) digitally zoomed in to demonstrate the fibrillary plaque structure (right), ( E ) Fluorescence intensity plot of THT stained plaque in ( D ) compared to a nearby stained cell.

Article Snippet: Thioflavin T (THT) , (MedChemExpress) , HY-D0218.

Techniques: Staining, Fluorescence

Journal: Acta histochemica

Article Title: Cytotoxicity induced by exposure to natural and synthetic tremolite asbestos: an in vitro pilot study.

doi: 10.1016/j.acthis.2012.04.004

Figure Lengend Snippet: Fig. 3. A549 cell viability measured by the MTT assay in control cultures and cultures exposed

Article Snippet: Cell cultures and treatments The biological effects of NAT and SAT were tested in A549 cells (CCL-185; American Type Culture Collection, Rockville, MD, 1 stoche U l i b p T ( l m s s b a s t f d m C y 2 b c 2 s s a r v f c u C o t c t p T 2 p f t c q t w s a J ( r d w F 1 d d E 02 A. Pugnaloni et al. / Acta Hi SA), a human malignant cell line with some features of alveoar epithelial cells.

Techniques: MTT Assay, Control

Journal: Acta histochemica

Article Title: Cytotoxicity induced by exposure to natural and synthetic tremolite asbestos: an in vitro pilot study.

doi: 10.1016/j.acthis.2012.04.004

Figure Lengend Snippet: Fig. 4. Toluidine blue staining. Semi-thin sections of control and NAT- and SAT-treated A549 cells at 24 and 48 h. Polygonal spread cells and sporadic mitotic elements (insert) were detected in control cultures; cellular nests and chains were more frequent at 48 h. Treated cultures showed few healthy isolated cells (H) and larger amounts of necrotic cells

Article Snippet: Cell cultures and treatments The biological effects of NAT and SAT were tested in A549 cells (CCL-185; American Type Culture Collection, Rockville, MD, 1 stoche U l i b p T ( l m s s b a s t f d m C y 2 b c 2 s s a r v f c u C o t c t p T 2 p f t c q t w s a J ( r d w F 1 d d E 02 A. Pugnaloni et al. / Acta Hi SA), a human malignant cell line with some features of alveoar epithelial cells.

Techniques: Staining, Control, Isolation

Journal: Acta histochemica

Article Title: Cytotoxicity induced by exposure to natural and synthetic tremolite asbestos: an in vitro pilot study.

doi: 10.1016/j.acthis.2012.04.004

Figure Lengend Snippet: Fig. 5. Time-lapse video microscopy recordings of control and NAT- and SAT-treated A549 cells at 24 h and 48 h. Loss of cell motility is clearly evident in treated cultures.

Article Snippet: Cell cultures and treatments The biological effects of NAT and SAT were tested in A549 cells (CCL-185; American Type Culture Collection, Rockville, MD, 1 stoche U l i b p T ( l m s s b a s t f d m C y 2 b c 2 s s a r v f c u C o t c t p T 2 p f t c q t w s a J ( r d w F 1 d d E 02 A. Pugnaloni et al. / Acta Hi SA), a human malignant cell line with some features of alveoar epithelial cells.

Techniques: Microscopy, Control

Journal: Acta histochemica

Article Title: Cytotoxicity induced by exposure to natural and synthetic tremolite asbestos: an in vitro pilot study.

doi: 10.1016/j.acthis.2012.04.004

Figure Lengend Snippet: Fig. 6. Fluorescence imaging of phalloidin staining for detection of F-actin at 24 h and 48 h. Control A549 cultures at 24 h and 48 h showing filamentous cytoskeletal actin (arrowheads). In NAT and SAT cells actin staining was irregular in the cytoplasm, with a more granular signal than in control cultures and a focal and spotted distribution. Insets: cells from the same cultures as Ctrl, NAT and SAT at higher magnification. Scale bars = 20 m; inserts = 10 m.

Article Snippet: Cell cultures and treatments The biological effects of NAT and SAT were tested in A549 cells (CCL-185; American Type Culture Collection, Rockville, MD, 1 stoche U l i b p T ( l m s s b a s t f d m C y 2 b c 2 s s a r v f c u C o t c t p T 2 p f t c q t w s a J ( r d w F 1 d d E 02 A. Pugnaloni et al. / Acta Hi SA), a human malignant cell line with some features of alveoar epithelial cells.

Techniques: Fluorescence, Imaging, Staining, Control

Journal: bioRxiv

Article Title: The Reissner fiber under tension in vivo shows dynamic interaction with ciliated cells contacting the cerebrospinal fluid

doi: 10.1101/2023.02.22.529498

Figure Lengend Snippet: (A1) Time-series standard deviation projection in the sagittal plane from 2 photon laser scanning microscope showing the signal from the Reissner fiber and CSF-cNs in the central canal of 3 dpf Tg ( sspo:sspo-GFP;pkd2l1:tagRFP;pkd2l1:GCaMP5G ) zebrafish larva before RF photoablation. (A2) Time-series standard deviation projection in the sagittal plane from 2 photon laser scanning microscope showing the signal from CSF-cNs after RF ablation performed by spiral scanning photoablation with an infrared pulsed laser tuned at 800 nm over 0.5 μm on the RF (see Methods). (B) ROI selection for ventral CSF-cNs to analyze activity before and after RF photoablation within the same cells (top). Example calcium activity traces normalized to baseline for each of the ROIs before (red) and after (black) RF photoablation over 75 s imaged at 3.45 Hz (see Methods). (C) Percentage of active ventral CSF-cNs (active is defined as having at least 1 calcium transient for the recording) before and after RF photoablation (109 cells total from 8 fish from 2 independent clutches; mean percent active before ablation = 27.72% ± 6.34% versus mean percent active after ablation = 10.59% ± 3.1%; paired two-tailed t-test: p < 0.05). (D) The ratio of active ventral CSF-cNs after RF photoablation to those active before RF photoablation, illustrating on average, a fraction (on average ± SEM: 45% ± 14%) of active ventral CSF-cNs before photoablation remain active after RF photoablation. The purple lines on the graph represent the mean and the error bars indicate the SEM. (E) Pie charts illustrating the number of events per ventral CSF-cN before and after RF photoablation (mean number of events in active cells before RF photoablation = 0.94 events / min versus mean number of events in active cells after RF photoablation = 0.87 events / min; paired two-tailed t-test: p < 0.005). *p < 0.05, ** p < 0.005 Scale bar is 20 μm (A1, A2), 10 μm (B).

Article Snippet: Two-photon spiral scanning ablations at 800 nm were performed with a two-photon laser scanning microscope (2p- vivo , Intelligent Imaging Innovations, Inc., Denver, Colorado, USA) equipped with a 20X objective (N.A.

Techniques: Standard Deviation, Laser-Scanning Microscopy, Selection, Activity Assay, Two Tailed Test

Journal: Journal of Biomedical Optics

Article Title: Red-shifted excitation enhances the sensitivity of red genetically encoded Ca 2+ indicator and enables crosstalk-free two-photon holographic optophysiology

doi: 10.1117/1.JBO.30.11.116003

Figure Lengend Snippet: Optical setup of 2P holographic microscope. Schematic diagram of the optical setup integrating holographic stimulation with 2P imaging (a), a photograph of a 2P holographic microscope (b), the calculation process for producing a 3D spot using an SLM (c), the MATLAB-based GUI for generating two-photon spots (d), and a representative fluorescent image (e) are shown.

Article Snippet: First, two-dimensional fluorescent images are obtained by a 2P scanning microscope system (C2 plus, Nikon) including control software (NIS Elements, Nikon) and a tunable femtosecond (fs) laser from 660 to 1300 nm at a pulse width of 100 fs and a repetition rate of 80 MHz (Chameleon Discovery NX, Coherent, Saxonburg, PA). shows a representative obtained fluorescent image.

Techniques: Microscopy, Imaging