Kubke Lab:/Notebook/Cranial nerve development/2010/11/24: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
m (→Satya Tutorial Day 1: adding embryo names and links) |
|||
Line 8: | Line 8: | ||
=General Entries= | =General Entries= | ||
=='''Satya Tutorial Day 1'''== | =='''Satya Tutorial Day 1'''== | ||
'''9.30 a.m. - 11.30 a.m.''': Firstly we discussed the different approaches for moulding the embryos. There were two incisions made to [[Kubke_Lab:Research/CND/Records/RC001|MH001]]. The reason behind doing this was to isolate the hindbrain region. During a later tutorial the decision was made not to separate the hindbrain and the head region, instead to make a single incision to separate the embryo at the level of the wind bud. [[Kubke_Lab:Research/CND/Records/RC001|RC001]] was moulded whole. Satya demonstrated the technique to create a mould for the tissue of interest using 'Tissue-Tek' OCT. Once the mould was created, the tissue was frozen for 5 minutes at -23 degrees Celsius. We created three moulds overall, one mould with a whole ST22 embryo ([[Kubke_Lab:Research/CND/Records/RC001|RC001]]). The second and third mould comprised of the ST28 embryo ([[Kubke_Lab:Research/CND/Records/RC001|MH001]]) with isolated sections of the head and hindbrain. Then we were briefed on the protocol and safety requirements associated with Cryostat cutting. Reuben then had practise in cutting his tissue using the cryostat and mounting the sections to slides. Malisha observed the cutting and helped with the labelling and mounting of slices onto slides. Once our histology tutorial is complete we hope to create a instructional page associated with cryostat specimen preparation and cutting. | '''9.30 a.m. - 11.30 a.m.''': Firstly we discussed the different approaches for moulding the embryos. There were two incisions made to [[Kubke_Lab:Research/CND/Records/RC001|MH001]]. | ||
{{Note|is it MH001 or RC001? --[[User:M Fabiana Kubke|MF Kubke]] 03:55, 3 February 2011 (EST)}} The reason behind doing this was to isolate the hindbrain region. During a later tutorial the decision was made not to separate the hindbrain and the head region, instead to make a single incision to separate the embryo at the level of the wind bud. [[Kubke_Lab:Research/CND/Records/RC001|RC001]] was moulded whole. Satya demonstrated the technique to create a mould for the tissue of interest using 'Tissue-Tek' OCT. Once the mould was created, the tissue was frozen for 5 minutes at -23 degrees Celsius. We created three moulds overall, one mould with a whole ST22 embryo ([[Kubke_Lab:Research/CND/Records/RC001|RC001]]). The second and third mould comprised of the ST28 embryo ([[Kubke_Lab:Research/CND/Records/RC001|MH001]]) with isolated sections of the head and hindbrain. Then we were briefed on the protocol and safety requirements associated with Cryostat cutting. Reuben then had practise in cutting his tissue using the cryostat and mounting the sections to slides. Malisha observed the cutting and helped with the labelling and mounting of slices onto slides. Once our histology tutorial is complete we hope to create a instructional page associated with cryostat specimen preparation and cutting. | |||
Revision as of 01:55, 3 February 2011
Cranial Nerve Development | <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page <html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> |
General EntriesSatya Tutorial Day 19.30 a.m. - 11.30 a.m.: Firstly we discussed the different approaches for moulding the embryos. There were two incisions made to MH001.
11.30 a.m. onwards: Completing benchmarks set by Fabiana. Cryostat protocol (Work in progress)Moulding
Tip: The best way to get the embryo flat on the mould is by having a coat of OCT on the mould before placing the embryo.
Personal Entries |