BrainPhys™ Imaging Optimized Medium

Serum-free neurophysiological basal medium for improved neuronal live imaging and function
BrainPhys™ Imaging Optimized Medium

Serum-free neurophysiological basal medium for improved neuronal live imaging and function

500 mL Bottle
Catalog # 05796
135 USD
Required Products
  1. NeuroCult™ SM1 Neuronal Supplement
    NeuroCult™ SM1 Neuronal Supplement

    Supplement (50X) for the serum-free culture of neurons

Overview

BrainPhys™ Imaging Optimized Medium is a serum-free neuronal basal medium (based on the BrainPhys™ formulation published by Cedric Bardy and Fred H. Gage) that is optimized for imaging applications.

Applications of BrainPhys™ Imaging Optimized Medium include live fluorescent imaging (calcium imaging and optogenetics) and neuronal culture. In addition to the removal of phenol red, the formulation has been modified to reduce background fluorescence and increase stability upon repeated exposure to light (M Zabolocki et al. Nat Comm, 2020).
Advantages
• No photoxicity for extended live imaging
• Reduced background autofluorescence in the 488 nm excitation channel
• Perform functional assays without changing media and shocking cells
• More representative of the brain's extracellular environment
• Improved neuronal function and a higher proportion of synaptically active neurons
• Rigorous raw material screening and quality control ensure minimal lot-to-lot variability
Components
  • BrainPhys™ Imaging Optimized Medium (Catalog #05796)
    • BrainPhys™ Imaging Optimized, 500 mL
Subtype
Specialized Media
Cell Type
Neural Cells, PSC-Derived, Neurons, Pluripotent Stem Cells
Species
Human, Mouse, Rat, Non-Human Primate, Other
Application
Cell Culture, Functional Assay, Immunofluorescence, Organoid Culture
Brand
BrainPhys
Area of Interest
Disease Modeling, Drug Discovery and Toxicity Testing, Neuroscience, Organoids
Formulation
Serum-Free, Chemically Defined

Scientific Resources

Product Documentation

Document Type Product Name Catalog # Lot # Language
Document Type
Product Information Sheet
Product Name
BrainPhys™ Imaging Optimized Medium
Catalog #
05796
Lot #
All
Language
English
Document Type
Safety Data Sheet
Product Name
BrainPhys™ Imaging Optimized Medium
Catalog #
05796
Lot #
All
Language
English

Educational Materials (15)

Brochure
STEMCELL Neural Product Portfolio
Brochure
Primary Neuronal Culture: Standardized Media and Reagents
Brochure
BrainPhys™ Neuronal Medium for Improved Neuronal Function
Brochure
NeuroFluor™ NeuO Selectively Label Live Neurons
Brochure
hPSC-Derived Neural Cell Research
Protocol
How to Generate AssemBloids™ from hPSC-Derived Dorsal and Ventral Forebrain Organoid Co-Cultures
Wallchart
Cell-Reprogramming Technology and Neuroscience
Video
BrainPhys™: A New Way to Culture Neurons
0:55
BrainPhys™: A New Way to Culture Neurons
Webinar
Building Brain Organoids and AssemBloids™ to Study Human Development and Disease
1:02:03
Building Brain Organoids and AssemBloids™ to Study Human Development and Disease
Webinar
The Road to Functional Human Neuronal Circuits in Vitro
56:27
The Road to Functional Human Neuronal Circuits in Vitro
Webinar
BrainPhys™ Medium Supports the Physiological Activity of Neuronal Tissue in vitro
27:19
BrainPhys™ Medium Supports the Physiological Activity of Neuronal Tissue in vitro
Scientific Poster
A New Culture Medium That Reduces Phototoxicity and Autofluorescence While Supporting Activity in Long-Term Primary Tissue- and hPSC-Derived Neurons
Scientific Poster
Determining the Metabolic Requirements of Electrically Active Rodent Primary Neurons in Long-Term Culture
Scientific Poster
Generation of a Glia-Neuron Co-Culture System Derived From Human Pluripotent Stem Cells
Scientific Poster
BrainPhys™ Neuronal Medium: A Medium Optimized to Support the Synaptic Activity of Neurons Derived from Human Pluripotent Stem Cells and Primary CNS Tissues
Load More Educational Materials

Product Applications

This product is designed for use in the following research area(s) as part of the highlighted workflow stage(s). Explore these workflows to learn more about the other products we offer to support each research area.

Data and Publications

Data

Experimental protocol schematic for live imaging of hPSC-derived neurons

Figure 1. Schematic for Live Imaging During Differentiation of hPSC-Derived Neurons

Neurons can be transitioned into BrainPhys™ IO with the relevant supplements and cultured for a maximum of 14 days.

Experimental protocol schematic for live imaging of primary neurons

Figure 2. Schematic for Live Imaging During Maturation of Primary Tissue-Derived Neurons

Neurons can be transitioned into BrainPhys™ IO with serum replacement supplement and cultured for a maximum of 14 days.

Culturing neurons in BrainPhys™ Imaging Optimized Medium reduces phototoxicity after blue light exposure and autofluorescence at a mean emission of 525 nm

Figure 3. BrainPhys™ IO Reduces Phototoxicity and Autofluorescence of Imaged Cells

Primary rat cortical neurons cultured in BrainPhys™ Imaging Optimized Medium (A) retain a healthy morphology after exposure to blue LED light for 12 hours. (B) Neurons labeled with live neuron dye NeuroFluor™ NeuO showed reduced background fluorescence at a mean emission of 525 nm, resulting in improved image contrast. This medium has enhanced performance under live imaging conditions compared to the original BrainPhys™ Neuronal Medium formulation, which provides superior long-term culture health, but shows (C) some disintegrated cell bodies and neurites (black arrows) and (D) autofluorescence under the same experimental conditions.

Figure 4. BrainPhys™ Imaging Optimized Medium Reduces Autofluorescence Relative to Standard Neural Media and Other Imaging-Specialized Media

The emission spectra across 400 - 700 nm of the light spectrum were captured as mean autofluorescence intensity from test and control (PBS) media (without cells) for the 375 (ultraviolet), 405 (violet), 488 (blue), and 532 nm (green) excitation wavelengths. Mean fluorescence intensity in PBS was subtracted from other media to normalize the data (n = 8 replicate wells, 3 independent experiments). BrainPhys™ Imaging Optimized (BPI) shows autofluorescence intensities similar to PBS and far lower intensities than are observed from standard neural media. Adapted from Zabolocki et al. 2020, Nature Communications, available under a Creative Commons 4.0 License.

Figure 5. Fluorescent Imaging in BrainPhys™ Imaging Optimized Medium Improves Signal-to-Background Ratios of 3D Neural Cultures

GFP-labeled ventral forebrain organoids were co-cultured and fused with unlabeled dorsal forebrain organoids for one week prior to live imaging in Forebrain Organoid Differentiation Medium from STEMdiff™ Dorsal Forebrain Organoid Differentiation Kit (right) or BrainPhys™ Imaging Optimized Medium (BPI, left). Interneuron migration can be visualized more clearly in BPI. Adapted from Zabolocki et al. 2020, Nature Communications, available under a Creative Commons 4.0 License.

Figure 6. BrainPhys™ Imaging Optimized Medium Supports Live Calcium Imaging of Human Neurons In Vitro

Intracellular Ca2+ changes in human PSC-derived neurons were measured with time-lapse imaging sequences of a Ca2+ sensor. Regions of interest (ROIs) were drawn on the cell soma to determine fluorescence intensity changes (ΔF/F0) over time. The same fields of view (FOVs) were imaged in artificial cerebrospinal fluid (ACSF), BrainPhys™ Imaging Optimized (BPI), Imaging Alternative 2, and Imaging Alternative 1 media. (A) Representative image of a neuronal population in BPI. White circles represent active ROIs, and fluorescent intensity represents intracellular Ca2+ levels. (B) Comparison of Ca2+ signals in BPI, Imaging Alternative 1 (Img Alt 1), and Imaging Alternative 2 (Img Alt 2) conditions show significant differences in the proportions of cells with Ca2+ spikes (two-tailed Mann-Whitney tests, mean ± SEM). Active cells with Ca2+ spikes in BPI (n = 243 cells), Imaging Alternative 2 (n = 39 cells), and Imaging Alternative 1 (n = 18 cells) were compared across seven FOVs from two coverslips. (C) Example traces from the same ROIs in different media. Adapted from Zabolocki et al. 2020, Nature Communications, available under a Creative Commons 4.0 License.

Figure 7. BrainPhys™ Imaging Optimized Medium Supports Long-Term Studies of Neuronal Activity Combined with Imaging-Based Characterization

Spontaneous firing rates of human neurons expressing a synapsin:ChETA-YFP optogene were recorded in BrainPhys™ (BP), BrainPhys™ Imaging Optimized (BPI), or Imaging Alternative 1 media with identical supplements using microelectrode arrays. Recordings were split into two groups (circles, n = 8 wells; or triangles, n = 7 wells). Both groups were switched from BP to BPI after 82 days, and only the triangle group was switched to Imaging Alternative 1 from day 89 - 95. From day 96, both groups were cultured in BP for one week before switching to BPI. Long-term firing is enhanced in BPI relative to Imaging Alternative 1, and mean firing rates (MFR, mean ± SEM) from both groups perform at baseline following the media change at day 96. Adapted from Zabolocki et al. 2020, Nature Communications, available under a Creative Commons 4.0 License.

Figure 8. BrainPhys™ Imaging Optimized Medium Supports Long-Term Studies of Human Neuronal Function In Vitro with Equivalent Performance to BrainPhys™ Medium

Multielectrode array (MEA) recordings from human pluripotent stem cell (hPSC)-derived neurons cultured in a 48-well MEA plate in BrainPhys™ (BP) for 9 weeks (n = 6 wells, 96 electrodes) or switched to BrainPhys™ Imaging Optimized (BPI) from BP during weeks 6 - 8 (n = 5 wells, 80 electrodes). (A) Percentage of active electrodes (>0.017Hz) and (B) mean firing rate (MFR) were both maintained during the period in BPI. Values are presented as mean ± SEM. Adapted from Zabolocki et al. 2020, Nature Communications, available under a Creative Commons 4.0 License.

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