Since the reaction of the PROT1 with the blood sample can be approximated with the Arrhenius equation, the reaction rate will be cut in half for every 10 degrees Celsius drop in temperature. Because of this, to ensure proper preservation of the blood sample, both the blood sample and the PROT should both be at room temperature before initiating the reaction. If the overall reaction temperature is too low then the sample will not be adequately fixed before being transferred to the freezer.
Here are a few additional tools to consider using which may improve consistency:
Sticker to make sure the PROT1 is at room temperature before using (should not be stored below room temperature but this would be visual confirmation that it is at room temperature):
https://www.zebra.com/us/en/products/environmental-sensors/ready-to-use-indicators/limitmarker-reversible.html
Use the same type of sticker on the Vacutainer to make sure the blood is at room temperature before adding to the PROT1
Data logger the size and shape of a Vacutainer:
https://www.motryx.com/
I don't know if the above can work down to -80C but the following one supposedly can:
https://www.logmore.com/products/data-loggers
This next one is disposable. It probably won't give you the amount of information you need right now, but the idea of a disposable temperature sensor could be helpful for the right application:
https://evigence.com/
Disposable exact transfer pipette (volume is too small for the 4ml volume discussed today but you may find this useful for the right project):
https://www.fishersci.com/shop/products/samco-exact-volume-transfer-pipettes/14754526
https://www.thermofisher.com/order/catalog/product/966NL
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