ESO wikiadmin: Created page with "{| class="wikitable" |+ !Materials |- |PBS pH 7.4 |- |4% PFA in PBS |- |10%, 20% and 30% sucrose solution in PBS |- |OCT Tissue-Tek |- |Histology moulds |- |Pre-coated glass slides |- |Dry ice |} === Procedure: === * Tissues/ anesthetized embryos are dissected and placed into 4% PFA solution overnight at 4° C * The next day the specimen is washed with PBS and transferred to a 10% sucrose solution for 3 h at 4° C (keep at 4°C until specimen sinks) * The sample is tra..."

2025-05-20T14:06:35Z

Created page with "{| class="wikitable" |+ !Materials |- |PBS pH 7.4 |- |4% PFA in PBS |- |10%, 20% and 30% sucrose solution in PBS |- |OCT Tissue-Tek |- |Histology moulds |- |Pre-coated glass slides |- |Dry ice |} === Procedure: === * Tissues/ anesthetized embryos are dissected and placed into 4% PFA solution overnight at 4° C * The next day the specimen is washed with PBS and transferred to a 10% sucrose solution for 3 h at 4° C (keep at 4°C until specimen sinks) * The sample is tra..."

New page

{| class="wikitable"
|+
!Materials
|-
|PBS pH 7.4
|-
|4% PFA in PBS
|-
|10%, 20% and 30% sucrose solution in PBS
|-
|OCT Tissue-Tek
|-
|Histology moulds
|-
|Pre-coated glass slides
|-
|Dry ice
|}

=== Procedure: ===

* Tissues/ anesthetized embryos are dissected and placed into 4% PFA solution overnight at 4° C
* The next day the specimen is washed with PBS and transferred to a 10% sucrose solution for 3 h at 4° C (keep at 4°C until specimen sinks)
* The sample is transferred to a 20% sucrose solution with subsequent incubation in 30% sucrose solution each for 3 hours at 4°C
* Incubate the sample on a rocking platform in a 1:1 mixture of OCT and 30% sucrose for 30 minutes
* Finally the sample is embedded in O.C.T Tissue-Tek using histology moulds, transferred to dry ice and stored at -80°C
* The sections are cut 10 µm thick in a cryostat at -20°C and transferred to pre- coated slides. The temperature of the cutting chamber is adjusted ±5°C according to the tissue specimen
* Sections are allowed to dry at RT for 2 hours and transferred to -20°C for storage

Tissue/embryo preparation fo cryosectioning - Revision history