brainphys imaging Search Results


brainphys tm imaging optimized medium  (STEMCELL Technologies Inc)
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STEMCELL Technologies Inc brainphys tm imaging optimized medium
Brainphys Tm Imaging Optimized Medium, supplied by STEMCELL Technologies Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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brainphys™ imaging  (STEMCELL Technologies Inc)
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STEMCELL Technologies Inc brainphys™ imaging
a The osmolality of <t>BrainPhys</t> Imaging (BPI) is similar to human cerebrospinal fluid (hCSF) (~305 mOsmol/kg). Neurobasal and NEUMO were significantly lower (210–220 mOsmol/kg) and DMEM/F12 and FluoroBrite higher (317–338 mOsmol/kg). Media osmolality data was collected from four replicates per condition. A total of three hCSF samples, each pooled from up to four subjects, were tested across three independent experiments ( n = 3). See also Supplementary Fig. . b – f Comparison of the optical properties of BPI with other basal media specialized for imaging (NEUMO, FluoroBrite), standard basal neuromedia (BrainPhys, BP; BrainPhys without phenol red, BP no PR; DMEM/F12; Neurobasal) and control media (phosphate-buffered saline, PBS; deionized water, H 2 O). b The absorbance spectra from 300 to 800 nm acquired from basal media alone (without cells), and after adding supplements required for the long-term culture of brain cells. Virtually all fluorophores used for cell imaging require light stimulation above 300 nm. c – f The mean autofluorescence intensities of basal media (without cells) across the entire visible spectrum. BPI shows autofluorescence intensities similar to PBS. c The emission spectra across 400–700 nm captured for the 375 (ultra-violet), 405 (violet), 488 (blue), and 532 nm (green) excitation wavelengths from test and control media. d – f Autofluorescence at 460, 520, and 590 nm emission wavelengths were measured following excitation at 355, 485, and 544 nm, respectively. Results were generated from eight replicate wells per medium ( n = 8) analyzed across three independent experiments. For normalization, the mean fluorescence intensity in PBS was subtracted from the other media. Data are presented as mean ± SEM. Significance determined via two-tailed nonparametric unpaired (Mann–Whitney) tests. ns, P > 0.05.
Brainphys™ Imaging, supplied by STEMCELL Technologies Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/brainphys™ imaging/product/STEMCELL Technologies Inc
Average 90 stars, based on 1 article reviews
brainphys™ imaging - by Bioz Stars, 2026-03
90/100 stars
  Buy from Supplier

Image Search Results


a The osmolality of BrainPhys Imaging (BPI) is similar to human cerebrospinal fluid (hCSF) (~305 mOsmol/kg). Neurobasal and NEUMO were significantly lower (210–220 mOsmol/kg) and DMEM/F12 and FluoroBrite higher (317–338 mOsmol/kg). Media osmolality data was collected from four replicates per condition. A total of three hCSF samples, each pooled from up to four subjects, were tested across three independent experiments ( n = 3). See also Supplementary Fig. . b – f Comparison of the optical properties of BPI with other basal media specialized for imaging (NEUMO, FluoroBrite), standard basal neuromedia (BrainPhys, BP; BrainPhys without phenol red, BP no PR; DMEM/F12; Neurobasal) and control media (phosphate-buffered saline, PBS; deionized water, H 2 O). b The absorbance spectra from 300 to 800 nm acquired from basal media alone (without cells), and after adding supplements required for the long-term culture of brain cells. Virtually all fluorophores used for cell imaging require light stimulation above 300 nm. c – f The mean autofluorescence intensities of basal media (without cells) across the entire visible spectrum. BPI shows autofluorescence intensities similar to PBS. c The emission spectra across 400–700 nm captured for the 375 (ultra-violet), 405 (violet), 488 (blue), and 532 nm (green) excitation wavelengths from test and control media. d – f Autofluorescence at 460, 520, and 590 nm emission wavelengths were measured following excitation at 355, 485, and 544 nm, respectively. Results were generated from eight replicate wells per medium ( n = 8) analyzed across three independent experiments. For normalization, the mean fluorescence intensity in PBS was subtracted from the other media. Data are presented as mean ± SEM. Significance determined via two-tailed nonparametric unpaired (Mann–Whitney) tests. ns, P > 0.05.

Journal: Nature Communications

Article Title: BrainPhys neuronal medium optimized for imaging and optogenetics in vitro

doi: 10.1038/s41467-020-19275-x

Figure Lengend Snippet: a The osmolality of BrainPhys Imaging (BPI) is similar to human cerebrospinal fluid (hCSF) (~305 mOsmol/kg). Neurobasal and NEUMO were significantly lower (210–220 mOsmol/kg) and DMEM/F12 and FluoroBrite higher (317–338 mOsmol/kg). Media osmolality data was collected from four replicates per condition. A total of three hCSF samples, each pooled from up to four subjects, were tested across three independent experiments ( n = 3). See also Supplementary Fig. . b – f Comparison of the optical properties of BPI with other basal media specialized for imaging (NEUMO, FluoroBrite), standard basal neuromedia (BrainPhys, BP; BrainPhys without phenol red, BP no PR; DMEM/F12; Neurobasal) and control media (phosphate-buffered saline, PBS; deionized water, H 2 O). b The absorbance spectra from 300 to 800 nm acquired from basal media alone (without cells), and after adding supplements required for the long-term culture of brain cells. Virtually all fluorophores used for cell imaging require light stimulation above 300 nm. c – f The mean autofluorescence intensities of basal media (without cells) across the entire visible spectrum. BPI shows autofluorescence intensities similar to PBS. c The emission spectra across 400–700 nm captured for the 375 (ultra-violet), 405 (violet), 488 (blue), and 532 nm (green) excitation wavelengths from test and control media. d – f Autofluorescence at 460, 520, and 590 nm emission wavelengths were measured following excitation at 355, 485, and 544 nm, respectively. Results were generated from eight replicate wells per medium ( n = 8) analyzed across three independent experiments. For normalization, the mean fluorescence intensity in PBS was subtracted from the other media. Data are presented as mean ± SEM. Significance determined via two-tailed nonparametric unpaired (Mann–Whitney) tests. ns, P > 0.05.

Article Snippet: On day 1 post-thaw, 500 µl of BrainPhys™ (Cat. No. 05790, STEMCELL Technologies) or BrainPhys™ Imaging (Cat. No. 05796, STEMCELL Technologies) supplemented with 1× NeuroCult™ SM1, 1× N2-A, 20 ng/ml BDNF (Cat. No. 78005, STEMCELL Technologies), 20 ng/ml GDNF (Cat. No. 78058, STEMCELL Technologies), 200 nM ascorbic acid (Cat. No. 72132, STEMCELL Technologies), 1 μg/ml laminin (Cat. No. L2020, Sigma) and 0.5 mM dibutyryl cyclic-AMP (Cat. No. 73882, STEMCELL Technologies) [complete BrainPhys™] was added per well.

Techniques: Imaging, Generated, Fluorescence, Two Tailed Test, MANN-WHITNEY

Fluorescent imaging in BrainPhys imaging (BPI) medium improves signal-to-background ratios. a – c Live human PSC-derived neurons expressing GFP and imaged in BPI, BrainPhys (BP) or artificial cerebrospinal fluid (ACSF) display higher mean GFP intensities at soma and neurite regions in BPI or ACSF relative to BP, with reduced background intensities. Data were analysed from the same cells (regions of interest, ROIs) in BP, BPI, or ACSF across nine field-of-views (FOVs) per condition; n = 293 ROIs analyzed at the soma, n = 469 ROIs analyzed at dendrites, n = 139 ROIs analyzed in background. b (left) Cumulative percentage (%) of pixels at background, soma and neurite regions in BP (gray), BPI (blue), and ACSF (black). Camera dark counts (without medium and cells) are labeled “dark”. b (right) Mean intensity values at background regions (top) in ACSF and BPI were significantly reduced relative to BP. Signal-to-background ratios at isolated neurite and soma regions were significantly higher in BPI and ACSF than BP. c Analysis of combined neurite and soma regions improved signal-to-background ratios in BPI and ACSF compared to BP. d Example images of brain assembloids representing a GFP-labeled ventral organoid fused with a nonlabeled dorsal organoid imaged in Organoid Maturation medium (right) or BPI (left). Increased visibility of interneuron migration is seen in BPI (bottom). e , f Representative primary cortical neurons stained and imaged live with Hoechst 33342, NeuroFluor NeuO, and fixed with β-III tubulin-Dylight 594. Images taken in both media are displayed with the following minimum/maximum intensity counts: 0/8500 (Hoechst), 0/3600 (NeuO), 1000/12,000 (β-III tubulin). Signal-to-background ratios were improved when imaging Hoechst in BPI ( n = 23 FOVs) compared to BP ( n = 24 FOVs). This was also seen for NeuO imaged in BPI ( n = 23 FOVs) relative to BP ( n = 22 FOVs). No significant differences were found when imaging β-III tubulin in either BPI ( n = 8 FOVs) or BP ( n = 9 FOVs). Images were collected from two biologically independent experiments (Hoechst/NeuO; β-III tubulin) with one well per condition. Data in b , c , f are presented as mean ± SEM. Significance determined via two-tailed nonparametric paired (Wilcoxon) tests ( b , c ) and nonparametric unpaired (Mann–Whitney) test ( f ). Nonsignificant P -values >0.05 are annotated with ns.

Journal: Nature Communications

Article Title: BrainPhys neuronal medium optimized for imaging and optogenetics in vitro

doi: 10.1038/s41467-020-19275-x

Figure Lengend Snippet: Fluorescent imaging in BrainPhys imaging (BPI) medium improves signal-to-background ratios. a – c Live human PSC-derived neurons expressing GFP and imaged in BPI, BrainPhys (BP) or artificial cerebrospinal fluid (ACSF) display higher mean GFP intensities at soma and neurite regions in BPI or ACSF relative to BP, with reduced background intensities. Data were analysed from the same cells (regions of interest, ROIs) in BP, BPI, or ACSF across nine field-of-views (FOVs) per condition; n = 293 ROIs analyzed at the soma, n = 469 ROIs analyzed at dendrites, n = 139 ROIs analyzed in background. b (left) Cumulative percentage (%) of pixels at background, soma and neurite regions in BP (gray), BPI (blue), and ACSF (black). Camera dark counts (without medium and cells) are labeled “dark”. b (right) Mean intensity values at background regions (top) in ACSF and BPI were significantly reduced relative to BP. Signal-to-background ratios at isolated neurite and soma regions were significantly higher in BPI and ACSF than BP. c Analysis of combined neurite and soma regions improved signal-to-background ratios in BPI and ACSF compared to BP. d Example images of brain assembloids representing a GFP-labeled ventral organoid fused with a nonlabeled dorsal organoid imaged in Organoid Maturation medium (right) or BPI (left). Increased visibility of interneuron migration is seen in BPI (bottom). e , f Representative primary cortical neurons stained and imaged live with Hoechst 33342, NeuroFluor NeuO, and fixed with β-III tubulin-Dylight 594. Images taken in both media are displayed with the following minimum/maximum intensity counts: 0/8500 (Hoechst), 0/3600 (NeuO), 1000/12,000 (β-III tubulin). Signal-to-background ratios were improved when imaging Hoechst in BPI ( n = 23 FOVs) compared to BP ( n = 24 FOVs). This was also seen for NeuO imaged in BPI ( n = 23 FOVs) relative to BP ( n = 22 FOVs). No significant differences were found when imaging β-III tubulin in either BPI ( n = 8 FOVs) or BP ( n = 9 FOVs). Images were collected from two biologically independent experiments (Hoechst/NeuO; β-III tubulin) with one well per condition. Data in b , c , f are presented as mean ± SEM. Significance determined via two-tailed nonparametric paired (Wilcoxon) tests ( b , c ) and nonparametric unpaired (Mann–Whitney) test ( f ). Nonsignificant P -values >0.05 are annotated with ns.

Article Snippet: On day 1 post-thaw, 500 µl of BrainPhys™ (Cat. No. 05790, STEMCELL Technologies) or BrainPhys™ Imaging (Cat. No. 05796, STEMCELL Technologies) supplemented with 1× NeuroCult™ SM1, 1× N2-A, 20 ng/ml BDNF (Cat. No. 78005, STEMCELL Technologies), 20 ng/ml GDNF (Cat. No. 78058, STEMCELL Technologies), 200 nM ascorbic acid (Cat. No. 72132, STEMCELL Technologies), 1 μg/ml laminin (Cat. No. L2020, Sigma) and 0.5 mM dibutyryl cyclic-AMP (Cat. No. 73882, STEMCELL Technologies) [complete BrainPhys™] was added per well.

Techniques: Imaging, Derivative Assay, Expressing, Labeling, Isolation, Migration, Staining, Two Tailed Test, MANN-WHITNEY

a – d Representative images of rat cortical primary neurons cultured in BrainPhys (BP) and BrainPhys Imaging (BPI) pre and post violet, blue and red LED exposure show dying neurons in BP (highlighted by white arrows). Each image is a representative cropped field-of-view (FOV). See also Supplementary Fig. . d Only BPI maintained neuronal viability post violet and blue LED exposure compared to BP, NEUMO, and BP without phenol red (BP no PR). Red LED exposure caused no apparent phototoxicity. Data were collected across 3–5 independent experiments, each including 75 field-of-views analyzed across three replicate wells. The number of neurons (per cm 2 ) post-light exposure were normalized to “no light” conditions. e , f Significant cytotoxicity in human midbrain (days 0–5: BPI and BP, n = 6 wells; days 6–7: BPI and BP, n = 5 wells) and cortical neurons (BP, n = 5; BPI, n = 4 wells) was seen when BPI and BP media (with supplements) were exposed to ambient tissue culture light for 24 h and fed to cells across 7 days. g Reactive oxygen species (H 2 O 2 ) in BP (without cells) was significantly increased following 24 h stimulation with ambient light, relative to BPI ( n = 12 wells per condition). Symbols represent cortical (triangle) or midbrain (square) supplements. h Ambient tissue culture light emission spectrum. i Blue LED light emission spectrum. j Relative H 2 O 2 levels of BPI and BP (without cells) after 24 h stimulation with blue LED light flashes over increasing power intensities ( n = 6 wells per condition). Data shown were normalized to “no light” conditions. See also Supplementary Fig. . k Mitochondrial health of primary rat cortical neurons in BPI and BP (supplemented with SM1) was assessed using a mitochondrial membrane potential indicator (JC-1). Following 1 h violet LED light exposure or 100 µM H 2 O 2 treatment, only neurons exposed to BPI maintained mitochondrial health. Data were collected across 3–5 independent experiments, across three biological replicates. Results are shown normalized to “no light’” conditions. See also Supplementary Fig. . Data in d – g , j – k are presented as mean ± SEM. Significance determined via two-tailed unpaired t -test ( d ), two-tailed nonparametric unpaired (Mann–Whitney) test ( e – g , k ) and one-sided sum-of-squares F -test ( j ). Nonsignificant P -values >0.05 are annotated with ns.

Journal: Nature Communications

Article Title: BrainPhys neuronal medium optimized for imaging and optogenetics in vitro

doi: 10.1038/s41467-020-19275-x

Figure Lengend Snippet: a – d Representative images of rat cortical primary neurons cultured in BrainPhys (BP) and BrainPhys Imaging (BPI) pre and post violet, blue and red LED exposure show dying neurons in BP (highlighted by white arrows). Each image is a representative cropped field-of-view (FOV). See also Supplementary Fig. . d Only BPI maintained neuronal viability post violet and blue LED exposure compared to BP, NEUMO, and BP without phenol red (BP no PR). Red LED exposure caused no apparent phototoxicity. Data were collected across 3–5 independent experiments, each including 75 field-of-views analyzed across three replicate wells. The number of neurons (per cm 2 ) post-light exposure were normalized to “no light” conditions. e , f Significant cytotoxicity in human midbrain (days 0–5: BPI and BP, n = 6 wells; days 6–7: BPI and BP, n = 5 wells) and cortical neurons (BP, n = 5; BPI, n = 4 wells) was seen when BPI and BP media (with supplements) were exposed to ambient tissue culture light for 24 h and fed to cells across 7 days. g Reactive oxygen species (H 2 O 2 ) in BP (without cells) was significantly increased following 24 h stimulation with ambient light, relative to BPI ( n = 12 wells per condition). Symbols represent cortical (triangle) or midbrain (square) supplements. h Ambient tissue culture light emission spectrum. i Blue LED light emission spectrum. j Relative H 2 O 2 levels of BPI and BP (without cells) after 24 h stimulation with blue LED light flashes over increasing power intensities ( n = 6 wells per condition). Data shown were normalized to “no light” conditions. See also Supplementary Fig. . k Mitochondrial health of primary rat cortical neurons in BPI and BP (supplemented with SM1) was assessed using a mitochondrial membrane potential indicator (JC-1). Following 1 h violet LED light exposure or 100 µM H 2 O 2 treatment, only neurons exposed to BPI maintained mitochondrial health. Data were collected across 3–5 independent experiments, across three biological replicates. Results are shown normalized to “no light’” conditions. See also Supplementary Fig. . Data in d – g , j – k are presented as mean ± SEM. Significance determined via two-tailed unpaired t -test ( d ), two-tailed nonparametric unpaired (Mann–Whitney) test ( e – g , k ) and one-sided sum-of-squares F -test ( j ). Nonsignificant P -values >0.05 are annotated with ns.

Article Snippet: On day 1 post-thaw, 500 µl of BrainPhys™ (Cat. No. 05790, STEMCELL Technologies) or BrainPhys™ Imaging (Cat. No. 05796, STEMCELL Technologies) supplemented with 1× NeuroCult™ SM1, 1× N2-A, 20 ng/ml BDNF (Cat. No. 78005, STEMCELL Technologies), 20 ng/ml GDNF (Cat. No. 78058, STEMCELL Technologies), 200 nM ascorbic acid (Cat. No. 72132, STEMCELL Technologies), 1 μg/ml laminin (Cat. No. L2020, Sigma) and 0.5 mM dibutyryl cyclic-AMP (Cat. No. 73882, STEMCELL Technologies) [complete BrainPhys™] was added per well.

Techniques: Cell Culture, Imaging, Two Tailed Test, MANN-WHITNEY

Single-cell patch-clamp recordings of iPSC-derived neurons matured in standard BrainPhys (BP)+ supplements medium for >12 weeks, and patch clamped in BrainPhys imaging (BPI) or artificial cerebrospinal fluid (ACSF). All patch-clamped neurons ( n = 35) included in the analysis were classified as “Type-5” and patched from a total of 18 coverslips. See also Supplementary Fig. . A subset of neurons ( n = 6) were recorded in both media. Each point on the graphs represents a single neuron. a (Top) Typical evoked action potential (AP) traces following a 500 ms depolarizing current step when patched in ACSF or BPI. a (Bottom) Current–voltage characteristics (I–V curve) with +5 mV current steps increments from rest at −70 mV. Voltage-dependent sodium (Nav) and potassium (Kv) current traces shown, respectively, below and above the x -axes. b Shows an example image of a single neuron filled with rhodamine following patch-clamp recordings. c Pooled data summarizing peak and threshold AP amplitudes (BPI, n = 16; ACSF, n = 15), and resting membrane potential values (BPI, n = 12; ACSF, n = 9). d – f Action potential properties (AP) were similar in BPI ( n = 16) and ACSF ( n = 15). d Rheobase values. e Peak afterhyperpolarization (AHP) amplitudes. f Maximum firing frequency of evoked APs (with amplitudes > −10 mV). g – i Current–voltage characteristics in BPI were similar to ACSF perfusate. g Peak NaV current amplitudes (BPI, n = 19; ACSF, n = 16). h , i Peak amplitudes of rapidly and slowly inactivating Kv currents (BPI, n = 7; ACSF, n = 13). j , k Membrane resistance (Rm) and capacitance values were similar between BPI ( n = 19) and ACSF ( n = 13). l – p Synaptic events mediated by AMPA receptors (excitatory postsynaptic currents, ePSCs) and GABAa receptors (inhibitory postsynaptic currents, iPSCs) were supported in both BPI ( n = 13) and ACSF ( n = 11) perfusates. l Typical spontaneous ePSC (top) and iPSC traces (bottom). m – p Average properties of spontaneous synaptic events recorded for 4 min. m Mean ePSC amplitudes. n Mean ePSC event frequency. o Max rise slopes of ePSCs. p Max decay slopes of ePSCs. Symbols in d – k, m – p represent human neurons tested first (triangles) or second (circle) in either medium. Data are presented as mean ± SEM. Significance determined via two-tailed nonparametric unpaired (Mann–Whitney) tests. ns, P > 0.05.

Journal: Nature Communications

Article Title: BrainPhys neuronal medium optimized for imaging and optogenetics in vitro

doi: 10.1038/s41467-020-19275-x

Figure Lengend Snippet: Single-cell patch-clamp recordings of iPSC-derived neurons matured in standard BrainPhys (BP)+ supplements medium for >12 weeks, and patch clamped in BrainPhys imaging (BPI) or artificial cerebrospinal fluid (ACSF). All patch-clamped neurons ( n = 35) included in the analysis were classified as “Type-5” and patched from a total of 18 coverslips. See also Supplementary Fig. . A subset of neurons ( n = 6) were recorded in both media. Each point on the graphs represents a single neuron. a (Top) Typical evoked action potential (AP) traces following a 500 ms depolarizing current step when patched in ACSF or BPI. a (Bottom) Current–voltage characteristics (I–V curve) with +5 mV current steps increments from rest at −70 mV. Voltage-dependent sodium (Nav) and potassium (Kv) current traces shown, respectively, below and above the x -axes. b Shows an example image of a single neuron filled with rhodamine following patch-clamp recordings. c Pooled data summarizing peak and threshold AP amplitudes (BPI, n = 16; ACSF, n = 15), and resting membrane potential values (BPI, n = 12; ACSF, n = 9). d – f Action potential properties (AP) were similar in BPI ( n = 16) and ACSF ( n = 15). d Rheobase values. e Peak afterhyperpolarization (AHP) amplitudes. f Maximum firing frequency of evoked APs (with amplitudes > −10 mV). g – i Current–voltage characteristics in BPI were similar to ACSF perfusate. g Peak NaV current amplitudes (BPI, n = 19; ACSF, n = 16). h , i Peak amplitudes of rapidly and slowly inactivating Kv currents (BPI, n = 7; ACSF, n = 13). j , k Membrane resistance (Rm) and capacitance values were similar between BPI ( n = 19) and ACSF ( n = 13). l – p Synaptic events mediated by AMPA receptors (excitatory postsynaptic currents, ePSCs) and GABAa receptors (inhibitory postsynaptic currents, iPSCs) were supported in both BPI ( n = 13) and ACSF ( n = 11) perfusates. l Typical spontaneous ePSC (top) and iPSC traces (bottom). m – p Average properties of spontaneous synaptic events recorded for 4 min. m Mean ePSC amplitudes. n Mean ePSC event frequency. o Max rise slopes of ePSCs. p Max decay slopes of ePSCs. Symbols in d – k, m – p represent human neurons tested first (triangles) or second (circle) in either medium. Data are presented as mean ± SEM. Significance determined via two-tailed nonparametric unpaired (Mann–Whitney) tests. ns, P > 0.05.

Article Snippet: On day 1 post-thaw, 500 µl of BrainPhys™ (Cat. No. 05790, STEMCELL Technologies) or BrainPhys™ Imaging (Cat. No. 05796, STEMCELL Technologies) supplemented with 1× NeuroCult™ SM1, 1× N2-A, 20 ng/ml BDNF (Cat. No. 78005, STEMCELL Technologies), 20 ng/ml GDNF (Cat. No. 78058, STEMCELL Technologies), 200 nM ascorbic acid (Cat. No. 72132, STEMCELL Technologies), 1 μg/ml laminin (Cat. No. L2020, Sigma) and 0.5 mM dibutyryl cyclic-AMP (Cat. No. 73882, STEMCELL Technologies) [complete BrainPhys™] was added per well.

Techniques: Patch Clamp, Derivative Assay, Imaging, Two Tailed Test, MANN-WHITNEY

a – f Midbrain human iPSC-derived neurons were matured for 100 days in BP before switching to standard BrainPhys (BP) or BrainPhys Imaging (BPI) for 2 h. The same midbrain supplements were added to all basal media. Network activity was compared on 48-well multielectrode array (MEA) plates over 7-min time intervals. Each value represents the average recorded across 16 electrodes per well, with a total of 18–21 wells per condition. a Example 2-min raster plots of neuronal network activity in BPI or BP show similar network activity (blue boxes) and spike histograms (gray, top). b – f Quantification of MEA network activity showed no significant differences between BPI and BP media for: b synchrony index ( n = 21 wells per condition), c number of network events per minute ( n = 21 wells per condition), d network interburst interval (IBI) coefficient of variation ( n = 18 wells per condition), e mean network burst duration (BPI, n = 18 wells; BP, n = 19 wells) and f mean IBI duration (BPI, n = 20 wells; BP, n = 21 wells). g , h Cortical and midbrain human iPSC-derived neurons were matured for 52 days in standard BP. Spontaneous firing rates were then compared using MEA recordings over 18 days in BP (black), BPI (blue) and artificial cerebrospinal fluid (ACSF; green). The same cortical or midbrain supplements were added to all three basal media. Spontaneous MEA activity was recorded for 10 min every 24 h across 16 electrodes per well for 29 wells of midbrain (9–10 wells per condition) and 20 wells of cortical (6–7 wells per condition) cultures. For both midbrain and cortical cultures, the spontaneous firing activity was similar in BP, BPI, and ACSF at first but progressively decreased in ACSF after several days. Data in b – h are presented as mean ± SEM. Significance determined via two-tailed nonparametric unpaired (Mann–Whitney) tests. Nonsignificant P -values >0.05 are annotated with ns.

Journal: Nature Communications

Article Title: BrainPhys neuronal medium optimized for imaging and optogenetics in vitro

doi: 10.1038/s41467-020-19275-x

Figure Lengend Snippet: a – f Midbrain human iPSC-derived neurons were matured for 100 days in BP before switching to standard BrainPhys (BP) or BrainPhys Imaging (BPI) for 2 h. The same midbrain supplements were added to all basal media. Network activity was compared on 48-well multielectrode array (MEA) plates over 7-min time intervals. Each value represents the average recorded across 16 electrodes per well, with a total of 18–21 wells per condition. a Example 2-min raster plots of neuronal network activity in BPI or BP show similar network activity (blue boxes) and spike histograms (gray, top). b – f Quantification of MEA network activity showed no significant differences between BPI and BP media for: b synchrony index ( n = 21 wells per condition), c number of network events per minute ( n = 21 wells per condition), d network interburst interval (IBI) coefficient of variation ( n = 18 wells per condition), e mean network burst duration (BPI, n = 18 wells; BP, n = 19 wells) and f mean IBI duration (BPI, n = 20 wells; BP, n = 21 wells). g , h Cortical and midbrain human iPSC-derived neurons were matured for 52 days in standard BP. Spontaneous firing rates were then compared using MEA recordings over 18 days in BP (black), BPI (blue) and artificial cerebrospinal fluid (ACSF; green). The same cortical or midbrain supplements were added to all three basal media. Spontaneous MEA activity was recorded for 10 min every 24 h across 16 electrodes per well for 29 wells of midbrain (9–10 wells per condition) and 20 wells of cortical (6–7 wells per condition) cultures. For both midbrain and cortical cultures, the spontaneous firing activity was similar in BP, BPI, and ACSF at first but progressively decreased in ACSF after several days. Data in b – h are presented as mean ± SEM. Significance determined via two-tailed nonparametric unpaired (Mann–Whitney) tests. Nonsignificant P -values >0.05 are annotated with ns.

Article Snippet: On day 1 post-thaw, 500 µl of BrainPhys™ (Cat. No. 05790, STEMCELL Technologies) or BrainPhys™ Imaging (Cat. No. 05796, STEMCELL Technologies) supplemented with 1× NeuroCult™ SM1, 1× N2-A, 20 ng/ml BDNF (Cat. No. 78005, STEMCELL Technologies), 20 ng/ml GDNF (Cat. No. 78058, STEMCELL Technologies), 200 nM ascorbic acid (Cat. No. 72132, STEMCELL Technologies), 1 μg/ml laminin (Cat. No. L2020, Sigma) and 0.5 mM dibutyryl cyclic-AMP (Cat. No. 73882, STEMCELL Technologies) [complete BrainPhys™] was added per well.

Techniques: Derivative Assay, Imaging, Activity Assay, Two Tailed Test, MANN-WHITNEY

a – e Optogenetics of human neurons with whole-cell patch-clamp recordings in BrainPhys Imaging (BPI), artificial cerebrospinal fluid (ACSF), and NEUMO. Each data point in the histograms represents a neuron. a Example images of neurons expressing synapsin:ChETA-YFP optogene and filled with rhodamine from the patch-pipette. b Whole-cell patch-clamp traces of neurons stimulated with 10 × 5 ms flashes of 0.4 mW blue light at 10 Hz in BPI or ACSF. Bottom of the graph shows corresponding spike raster plots. c Neurons patched in ACSF or BPI and stimulated with identical light parameters in b show no significant difference across ChR2 currents (BPI, ACSF; n = 7), action potential (AP) success rates (BPI, n = 7; ACSF, n = 8) and max AP amplitudes (BPI, ACSF; n = 8). Symbols represent neurons tested first (triangles) or second (circles) in either medium. d Optogenetic responses from the same patch-clamped neuron under 0.1 mW of blue light in BPI, NEUMO, and BPI (recovery) show optimal optogenetic control in BPI. e Quantification of patched neurons ( n = 22 across four coverslips) recorded in BPI or NEUMO, stimulated with 10 × 5 ms flashes of blue light at 10 Hz with increasing power. f – i Optogenetically evoked and spontaneous firing rates of human neurons expressing synapsin:ChETA-YFP recorded in BP, BPI or NEUMO (with identical supplements) using multielectrode arrays (MEAs). Neurons were cultured in standard BP for 82 days before transitioning to “test” media. f , g Recordings were split into two groups: “circles’” (8 wells) and “triangles” (7 wells). Both groups were changed to BPI at day 82; only “triangles” switched to NEUMO at days 89–95. From day 96, both groups were cultured in BP for one week before switching to BPI. h , i Optogenetic responses of human neurons on MEAs in NEUMO ( n = 112 electrodes) or BPI ( n = 128 electrodes) when stimulated with blue light (Lumos) at increasing intensities. The cumulative number of spikes was summed in binning windows; h 27.5 ms bins; i 2 s bins from first light stimulus over three sweeps. Data in h were plotted with quadratic non-linear curves. Values in c , e , g , i are presented as mean ± SEM. Significance determined via two-tailed nonparametric unpaired (Mann–Whitney) tests. ns, P > 0.05.

Journal: Nature Communications

Article Title: BrainPhys neuronal medium optimized for imaging and optogenetics in vitro

doi: 10.1038/s41467-020-19275-x

Figure Lengend Snippet: a – e Optogenetics of human neurons with whole-cell patch-clamp recordings in BrainPhys Imaging (BPI), artificial cerebrospinal fluid (ACSF), and NEUMO. Each data point in the histograms represents a neuron. a Example images of neurons expressing synapsin:ChETA-YFP optogene and filled with rhodamine from the patch-pipette. b Whole-cell patch-clamp traces of neurons stimulated with 10 × 5 ms flashes of 0.4 mW blue light at 10 Hz in BPI or ACSF. Bottom of the graph shows corresponding spike raster plots. c Neurons patched in ACSF or BPI and stimulated with identical light parameters in b show no significant difference across ChR2 currents (BPI, ACSF; n = 7), action potential (AP) success rates (BPI, n = 7; ACSF, n = 8) and max AP amplitudes (BPI, ACSF; n = 8). Symbols represent neurons tested first (triangles) or second (circles) in either medium. d Optogenetic responses from the same patch-clamped neuron under 0.1 mW of blue light in BPI, NEUMO, and BPI (recovery) show optimal optogenetic control in BPI. e Quantification of patched neurons ( n = 22 across four coverslips) recorded in BPI or NEUMO, stimulated with 10 × 5 ms flashes of blue light at 10 Hz with increasing power. f – i Optogenetically evoked and spontaneous firing rates of human neurons expressing synapsin:ChETA-YFP recorded in BP, BPI or NEUMO (with identical supplements) using multielectrode arrays (MEAs). Neurons were cultured in standard BP for 82 days before transitioning to “test” media. f , g Recordings were split into two groups: “circles’” (8 wells) and “triangles” (7 wells). Both groups were changed to BPI at day 82; only “triangles” switched to NEUMO at days 89–95. From day 96, both groups were cultured in BP for one week before switching to BPI. h , i Optogenetic responses of human neurons on MEAs in NEUMO ( n = 112 electrodes) or BPI ( n = 128 electrodes) when stimulated with blue light (Lumos) at increasing intensities. The cumulative number of spikes was summed in binning windows; h 27.5 ms bins; i 2 s bins from first light stimulus over three sweeps. Data in h were plotted with quadratic non-linear curves. Values in c , e , g , i are presented as mean ± SEM. Significance determined via two-tailed nonparametric unpaired (Mann–Whitney) tests. ns, P > 0.05.

Article Snippet: On day 1 post-thaw, 500 µl of BrainPhys™ (Cat. No. 05790, STEMCELL Technologies) or BrainPhys™ Imaging (Cat. No. 05796, STEMCELL Technologies) supplemented with 1× NeuroCult™ SM1, 1× N2-A, 20 ng/ml BDNF (Cat. No. 78005, STEMCELL Technologies), 20 ng/ml GDNF (Cat. No. 78058, STEMCELL Technologies), 200 nM ascorbic acid (Cat. No. 72132, STEMCELL Technologies), 1 μg/ml laminin (Cat. No. L2020, Sigma) and 0.5 mM dibutyryl cyclic-AMP (Cat. No. 73882, STEMCELL Technologies) [complete BrainPhys™] was added per well.

Techniques: Optogenetics, Patch Clamp, Imaging, Expressing, Transferring, Cell Culture, Two Tailed Test, MANN-WHITNEY

a (left) Shows representative images of 14 days-in-vitro (DIV) rat cortical primary neurons and hPSC-derived neurons matured in BrainPhys (BP) and BrainPhys Imaging (BPI) exhibiting healthy neuron morphology. a (right) Representative immunocytochemistry images showing that 21 DIV primary rat cortical neurons and 14 DIV hPSC-derived neurons characterized by MAP2 expression (magenta) matured in BP or BPI also display an appropriate expression of synaptic marker Synapsin1 (green). b Quantification of the number of neurons per cm 2 as represented in a shows that BPI supports equivalent neuronal survival relative to BP for 21 DIV rat cortical primary neurons ( n = 11) and 14–21 DIV hPSC-derived neurons ( n = 9). Values represent mean ± SEM. Significance determined using two-tailed nonparametric unpaired (Mann–Whitney) tests; ns P > 0.05. c Multielectrode array (MEA) recordings from hPSC-derived neurons cultured in a 48-well MEA plate in BP for 9 weeks ( n = 6 wells, 96 electrodes) or switched to BPI from BP during weeks 6–8 ( n = 5 wells, 80 electrodes) represent the spontaneous mean firing rates and percentage of active electrodes (>0.017 Hz) for each data set. The percentage of active electrodes and mean firing rates were both maintained during the period in BPI. Values are presented as mean ± SEM.

Journal: Nature Communications

Article Title: BrainPhys neuronal medium optimized for imaging and optogenetics in vitro

doi: 10.1038/s41467-020-19275-x

Figure Lengend Snippet: a (left) Shows representative images of 14 days-in-vitro (DIV) rat cortical primary neurons and hPSC-derived neurons matured in BrainPhys (BP) and BrainPhys Imaging (BPI) exhibiting healthy neuron morphology. a (right) Representative immunocytochemistry images showing that 21 DIV primary rat cortical neurons and 14 DIV hPSC-derived neurons characterized by MAP2 expression (magenta) matured in BP or BPI also display an appropriate expression of synaptic marker Synapsin1 (green). b Quantification of the number of neurons per cm 2 as represented in a shows that BPI supports equivalent neuronal survival relative to BP for 21 DIV rat cortical primary neurons ( n = 11) and 14–21 DIV hPSC-derived neurons ( n = 9). Values represent mean ± SEM. Significance determined using two-tailed nonparametric unpaired (Mann–Whitney) tests; ns P > 0.05. c Multielectrode array (MEA) recordings from hPSC-derived neurons cultured in a 48-well MEA plate in BP for 9 weeks ( n = 6 wells, 96 electrodes) or switched to BPI from BP during weeks 6–8 ( n = 5 wells, 80 electrodes) represent the spontaneous mean firing rates and percentage of active electrodes (>0.017 Hz) for each data set. The percentage of active electrodes and mean firing rates were both maintained during the period in BPI. Values are presented as mean ± SEM.

Article Snippet: On day 1 post-thaw, 500 µl of BrainPhys™ (Cat. No. 05790, STEMCELL Technologies) or BrainPhys™ Imaging (Cat. No. 05796, STEMCELL Technologies) supplemented with 1× NeuroCult™ SM1, 1× N2-A, 20 ng/ml BDNF (Cat. No. 78005, STEMCELL Technologies), 20 ng/ml GDNF (Cat. No. 78058, STEMCELL Technologies), 200 nM ascorbic acid (Cat. No. 72132, STEMCELL Technologies), 1 μg/ml laminin (Cat. No. L2020, Sigma) and 0.5 mM dibutyryl cyclic-AMP (Cat. No. 73882, STEMCELL Technologies) [complete BrainPhys™] was added per well.

Techniques: In Vitro, Derivative Assay, Imaging, Immunocytochemistry, Expressing, Marker, Two Tailed Test, MANN-WHITNEY, Cell Culture