I think very little is known about this topic and also how to find out, unfixed cells of both kinds have to be measured first. If tissue samples, not single cells are fixed its likely that surface cells react different from cells situated inside the sample. What happens is certainly temperature dependent, also wether there are fluid currents among the cells think its very very difficult the make comparable experiments.

Not really an answer, but as you might have seen too as a starter:

the thread starting in Jan 2013:

https://www.researchgate.net/post/Measuring_adipocyte_cell_size_from_frozen_tissue_or_tissue_fixed_in_formalin_ethanol?exp_tc=tprc

If I can find a reference out of the old literature I will post it here. Best wishes and regards,

Wolfgang

I think very little is known about this topic and also how to find out, unfixed cells of both kinds have to be measured first. If tissue samples, not single cells are fixed its likely that surface cells react different from cells situated inside the sample. What happens is certainly temperature dependent, also wether there are fluid currents among the cells think its very very difficult the make comparable experiments.

Hi Martin,

I think the key to your question lies in the content of water, as shrinkage after formalin embedding is an effect of dehydration. Water content depends on age, gender, general nutritional status and so on - you might be able to contact this group (poster in German, tile means 'physical characteristics of adipose tissue - comparative analysis regarding adults and children as basis for the development of human models')

www.rechtsmedizin.med.uni-muenchen.de/service/.../sdt2010_berlin_fett.pdf

Formalin is an additive fixative.It croslink the proteins causing cell shrinkage.The shrinkage of cells is not found to be different based on the maturation level of cells.

Dear all,

first of all I must admit/can confirm that it really is obviously impossible to search specifically for articles dealing with the matter posted by Martin Rune Christensen. I have found two or three articles (one of them related to measurement of shrinkage during processing of cryosections (see below). I think that Carl Morton Laane is right in saying that any comparison of "tissular events" during processing have to be compared to measurments on native tissue. (also d'accord with his following views). This might be tricky if not impossible with the originally question. This "question" a little bit is confusing since the "application" of 4% formaldehyde as a fixative will not exert much volume changes (at least when applied properly and not to short or to long, temperature also important). All changes in volume IMHO will be initiated by the dehydration up the way into the embedding medium (be it paraffin or resin) and after that one is faced also with alterations of (2-Dim) measures of diameter, "volume" of section of fat cells/fat droplets/vacuoles during sectioning and even staining (provided that fat/lipid content of your "cells" has been preserved properly....i.e has been "fixed" in such a way that you at least can discriminate between and cells/cell parts).

FA is a NON-COAGULANT Fixative (which can be eliminated after very short fixation by washing in water or buffer).

@K.Sujathan: usually it is thought that the main component inducing shrinkage of tissues is connective tissue matrix (collagen etc.), not the protein content of fairly small adipocytes with only minor cytoplasm.

@Patricia Kohlhof: it might be that the group in Munich has answers. If someone had made contact meanwhile it would be intersting to learn what they have found out. in : fat vacuoles (I am not confident about!) may contain perhaps only a very very small amount of water which doesn't count for shrinkage during fixation. It is for sure the elution of fat/lipid material during dehydration and embedding procedures.

A paper which comes to my mind (reading FA-fixation basics about perhaps, unfortunately NOT for Fat cells but for LIVER) is:

at: http://decapoda.nhm.org/pdfs/1505/1505.pdf

The Journal of Histochemistry and Cytochemistry Vol. 33, No. 8, pp. 845-853, 1985

CECIL H. FOX, FRANK B. JOHNSON, JOHN WHITING, and PETER P. ROLLER

where in the section: < Shrinkage of Tissues by Formaldehyde > one can read:

- in their CONCLUSION you'll find:

There cited also as Ref. 16: might be another article worth to be read:

Jones D: Reactions of aldehydes with unsaturated fatty acids during histological fixation. In: Fixation in Histochemistry. Edited by PJ Stoward. Chapman and Hall, London, 1973.

The second paper I found was:

http://www.jlr.org/content/12/5/521.full.pdf

JOURNAL OF LIPID RESEARCH, VOL 12, 1971 521-530

L. SJÖSTRÖM, P. BJORNTORP, and J. VRANA.

Last but not least there is considerable doubt about (buffered) formaldehyde (alone) being the fixative of choice for such a task, remembering:

(cf. other threads / postings concerning that matter in ResearchGate/Topic Histology, cf.: http://www.researchgate.net/topic/Histology/post/Is_there_any_suitable_staining_method_protocol_for_lipids_in_paraffin_sections? )

Would be interested to see whether someone out in the wild has a practical solution for the technical problem of Martin Rune Christensen....

Best regards, Wolfgang

Thank you all for your comments - especially Wolfgang for your thorough answer. I have started testing af setup based on Sjöström, Björntorps and Vránas article (JOURNAL OF LIPID RESEARCH, VOL 12, 1971 521-530

L. SJÖSTRÖM, P. BJORNTORP, and J. VRANA.).

The main problem is that I am dealing with forensic autopsy material with different post mortem intervals of the tissue (I am excluding the deceased of moderate to severe putrefication). So Carl Morten Laane are indeed right that comparable experiments "will be very very difficult".

However I am grateful for the insights and thoughts you have given regarding fixation and shrinkage of fatty tissue.

@Patricia Kohlhof: I might just try and contact the group in Munich.

Dear Martin, you're welcome!

your using forensic material certainly will be another challenge, I know (and certainly not only for adipose tissue (;-( )

Best regards and wishes, Wolfgang

To the Crew of ResearchGate:

IMHO, the statistics of this original question for the READS are quite wrong:

as of 2018-08-11,@09:08 p.m. the "new" "official count" scores:

0 Recommendations - 7 Answers - 30 Reads - 14 Followers

By the date: ! 2014-10-08 !, ca. 20:38/08:38 p.m. ,

I personally noted for the question stats:

0 (UPVOTES) Recomm / 0(downvotes) · 7 Answers · 444 Views 13 followers

Where that decrease in view might come?? I know that nobody wants to lie...so I see: numbers can be manipulated (by accident? by the system? .... numbers lost?) ! (;-() W.M.

I agree with prof. Wolfgang H. Miss.

Interest questio.

Good and interesting question.

Interesting with the numbers of views for the question... Must be a system error. I have only recently broke 500 reads of which only ca. 100 were question reads.

As a follow up on the original question: I did not manage to find or determine the degree of adipocyte shrinkage. Using stereology, I measured the adipocyte profile diameter of S100 stained adipose tissue slides from the epicardium, greater omentum, abdominal subcutaneous AT and gluteal subcutaneous AT.

I discussed the matter with Professor in Stochastic Geometry and Stereology, Jens Nyengaard, and we decided that the relative shrinkage between the four AT compartments would be similar. Besides, as many studies investigating adipocyte morphology use formalin fixed and paraffin embedded tissue, comparing the conclusions of the studies is appropriate. However, directly comparing cell profile diameter between studies may be risky due to differences in staining and measuring techniques.

I found some studies looking into adipocyte morphology:

Article Comparative analysis of three human adipocyte size measureme...

Article Pathophysiology of Human Visceral Obesity: An Update

And one comparing adipocyte shrinkage in paraffin and plastic embedded tissue:

https://www.obesityresearchclinicalpractice.com/article/S1871-403X(12)00021-X/fulltext