By H. William Detrich III, Monte Westerfield, Leonard Zon
The Zebrafish: mobile and Developmental Biology, half A mobile Biology, is the newest variation within the Methods in mobile Biology sequence that appears at equipment for studying mobile and developmental biology of zebrafish. Chapters hide such issues as mobile biology and developmental and neural biology.
- Covers sections on version structures and sensible experiences, imaging-based ways, and rising studies
- Written by way of specialists within the field
- Contains state of the art fabric with regards to developmental biology in zebrafish
- New half version of this significant volume
Read or Download The Zebrafish Cellular and Developmental Biology, Part A Cellular Biology PDF
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Additional resources for The Zebrafish Cellular and Developmental Biology, Part A Cellular Biology
Cells of interest (prospective progenitors) should be isolated, counted, and resuspended in 100 mL of ZKS/ZEST medium and added to complete methylcellulose, along with cytokines, small molecules, or other agents to be investigated. 3 mg/mL recombinant zebrafish Gcsf to methylcellulose medium. 1 mg/mL recombinant zebrafish Epo to methylcellulose medium. 4.
For analysis, remove hematopoietic cells from culture at desired time points as described earlier and perform FACS. CFSE is read in the FL-1 channel (on FACS Caliber) or with most GFP filters (FACS Aria I and II, LSR-II), and will decrease in fluorescence intensity as cells divide. Compare divisions to Day 0 time point with FloJo software (TreeStar, Ashland, OR). We recommend using the BD LSR-II flow cytometer, as the different scatter profile of mature cells is easily distinguished and directly comparable to profiles shown in Fig.
Allow the mixture to cool to room temperature before adding 2x L-15 medium powder (Mediatech). Then, adjust the weight of the methylcellulose mixture to 1000 g with sterile water. Methylcellulose should be allowed to thicken at 4 C overnight before being aliquoted and stored at À20 C. 5 mL of complete methylcellulose to sterile round-bottom 14-mL tubes (Becton Dickinson) with 5-mL syringes and 16gauge needles for each condition. Cells of interest (prospective progenitors) should be isolated, counted, and resuspended in 100 mL of ZKS/ZEST medium and added to complete methylcellulose, along with cytokines, small molecules, or other agents to be investigated.
The Zebrafish Cellular and Developmental Biology, Part A Cellular Biology by H. William Detrich III, Monte Westerfield, Leonard Zon