For some experiments, it is required to perform a viability assessment and a sequential biopsy collection at a later time point (T1) than the initial clean collection (T0).

Experimental Preparation

Carry out the clean collection at T0 as described in the Clean Biopsy Collection section.

For the viability and sequential biopsy, ensure that you continue to observe the RNA handling guidelines are observed as described in the RNA Handling Guidelines in Clean Biopsy Collection. 

Experimental Procedure

For the viability and sequential biopsy, ensure that the sample storage guidelines are observed as described in Sample storage section in Clean Biopsy Collection. Samples should remain stored in the fridge until all samples have been collected (until after T1). The samples may then be transferred to the freezer or processed immediately.

Following the T1 collection, apply the selected compound to the well of biopsied cells. Switch on the heating and the CO2 in the OMNIUM.

Set up and execute an observation workflow as previously described in the Observation section in Feasibility of Biopsy Viability. 

Tip: If the observation workflow is to be run overnight, ensure that the UV in the OMNIUM is also switched off.

When the observation workflow is finished, run the viability assessment (as described in the Viability Assessment section in Feasibility of Biopsy Viability). Following the viability assessment, track the cells using the tracking tool (as described in Cell Tracking Tool section in Feasibility of Biopsy Viability).

Tip: When preparing the collection buffer, ensure that there is enough for collections at both T0 and T1.

For each viable, relocatable cell, carry out a clean biopsy collection as described in the Biopsies section in Clean Biopsy Collection).

Out-of-OMNIUM controls only need to be prepared once per experimental batch. Therefore, if the controls have already been prepared at T0, there is no need to replicate at T1.