Improve Research Reproducibility A Bio-protocol resource

number of samples and the speed of centrifugation

Zhou Yue Nov 11, 2023

Protocol: Isolation and Quantification of Metabolite Levels in Murine Tumor Interstitial Fluid by LC/MS

As tumor samples obtained from clinic are generally small and there is concern that the extracted metabolites are not enough to detect and analyse, is it possible to pool the interstitial fluid extracted from several tumors as a replicate?

Besides, I am also confused about the sp Read more Read less more less

29 Views

Answer

Recommend

1 answer

Sort by: most helpful / most recent

Alexander Muir Author Answered Nov 13, 2023

Ben May Department for Cancer Research, University of Chicago, Chicago

Pooled analysis has been done in other studies where sample volume has been limiting (PMID: 36991126, 34179825).

Regarding centrifugation speed, other studies have successfully used higher centrifugation speeds (~400g) without apparent intracellular contamination of TIF, including our own (PMID: 37254839). So, one can likely use higher centrifugation speeds. However, with any TIF collection, and especially collections at higher centrifugation speeds, I would recommend performing downstream analysis (e.g. LDH analysis and other analyses as recommended in PMID: 21541282) to ensure the samples collected are pure extracellular fluid and not contaminated with intracellular fluid from cells lysed during the TIF extraction.

1 helpful

0 unhelpful

1 comments down

My answer

Write your answer...

References (optional)

add Add more

Post an Answer