We are performing serial kidney biopsies on patients with diabetic kidney disease to identify the underlying mechanisms of disease. We recently found tissue samples stored in formalin from the first kidney biopsy that we performed more than 7 years ago on 22 of these participants. Measurement of these samples will greatly enhance the value of the data we have in these participants, since we only had data from other tissue specimens that had not been stored in formalin. For scientific validity, all measurements on these samples need to be performed by the same laboratory at the University of Minnesota that performed all other morphometric measurements in this cohort.
The work to be done for each of 22 light microscopy formalin fixed samples would include:
1. embedding in paraffin
2. serial sectioning through the blocks at 5 microns, - 200 sections/biopsy
3. PAS staining of every 5th section
4. Digital imaging of cortex in the stained sections
5. Point the digitized images for counting for all of tubular and interstitial parameters including:
No Scar Vv(lnt/cortex) =fraction of cortex that is interstitial; no scar tissue
No Scar Vv(PT/cortex) = fraction of cortex that is proximal tubules; no scar tissue
No Scar Vv(DT/cortex) =fraction of cortex that is distal & collecting tubules; no scar tissue
No Scar Vv(AT/cortex) =fraction of cortex that is atrophic tubules; no scar tissue
No Scar Vv(TT/cortex) =fraction of cortex that is Total Tubules (Proximal+Distal+Atrophic); no
scar tissue
No Scar Vv(ATITT) =fraction of Total Tubules that are Atrophic Tubules; no scar tissue
Vv(lnt/Scar-cortex) =fraction of scar-cortex that is interstitium
Vv(Total Int/Total Cortex)= fraction of Total Cortex that is Total lnterstitium
Vv(Total AT/Total Cortex)= fraction of Total Cortex that is Total Atrophic Tubules
Vv(Scar/Total Cortex)= fraction of Total Cortex that is Scar-cortex
6. Glomerular volume by Cavalieri
6. Mauer evaluation of biopsies for %GS, %FSGS, %atrophic gloms, index of arteriolar
hyalinosis, and descriptions of patterns of injury such as KW nodules, exudative glomerular
lesions and interstitial inflammation.
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