製品の特長

BrainPhys™ Imaging Optimized Mediumをもちいて、ニューロンの生細胞イメージングと機能を改善できます

• 長時間のライブイメージングの際にも光毒性を抑えます
• 488 nm励起チャネルにおける自己蛍光バックグラウンドを低減します
• 培地交換で細胞に衝撃を与えることなく機能アッセイを実施できます
• 脳の細胞外環境をよりよく再現できます
• ニューロン機能を改善し、シナプス活性のあるニューロンの割合を増やします
• 厳格な原料スクリーニングと品質管理により、ロット間のばらつきを最小化します

生細胞イメージングの流れ

hPSC由来ニューロンの分化過程におけるライブイメージング

ニューロンは、関連するサプリメントを加えたBrainPhys Imaging Optimized Mediumに移行し、最大14日間培養できます。

プライマリー組織由来ニューロンの成熟過程におけるライブイメージング

ニューロンは、血清代替サプリメントを加えたBrainPhys Imaging Optimized Mediumに移行し、最大14日間培養できます。

データ紹介

イメージング時の光毒性と自家蛍光を低減します

BrainPhys Imaging Optimized Medium（A）で培養された初代ラット皮質ニューロンは、青色LEDライトに12時間さらされた後も健康な形態を維持します。 （B）ライブニューロン色素NeuroFluor™ NeuOで標識されたニューロンは、525 nmの露光においてもバックグラウンド蛍光が減少し、画像コントラストが向上しました。この培地は元のBrainPhys Neuronal Mediumと比べてライブイメージング条件下での性能が向上しています。
BrainPhys Neuronal Mediumは、長期培養において細胞を優れた健康状態に維持することができますが、同じ実験条件では（C）いくつかの崩壊した細胞体と神経突起（黒い矢印）および（D）自家蛍光が見られます。

標準的な神経用培地やイメージング用培地と比べて自家蛍光を低減します

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.

3次元神経培養の蛍光イメージングで、バックグラウンドに対するシグナル強度比を改善します

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.

ヒト神経のin vitroでのライブカルシウムイメージングをサポートします

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.

神経活動の長期研究をサポートします

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.

ヒト神経機能の長期研究をBrainPhys™と同様にサポートします

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.