Histological and Quantitative Analysis of Prostate Cancer Xenografts

Intra-femoral tumors were fixed in 10% formalin, decalcified with 10% EDTA, embedded in OCT and frozen in isopentane/dry ice bath for cryosections or paraffin embedded as described in Raheem et al. [14 (link)]. Subcutaneous tumors were fresh frozen in OCT or 10% formalin-fixed and embedded in paraffin at the time of harvest. IF and SC sections were H&E and immunostained with anti-PSA, 1:500 (DAKO) [14 (link)]. Images were captured using the Aperio ScanScope and Keyence digital microscope. Quantitation of PSA staining in FFPE sections was performed using The Spectrum Analysis algorithm package and ImageScope Analysis software as described in Woo et al. [18 (link)] with the modifications that five fields of view were randomly selected in Aperio ScanScope digital images at 20 X magnification in each of three different tumors in each treatment group. Total anti-PSA stained area/total analysis area was calculated for each field of view.

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Godebu E., Muldong M., Strasner A., Wu C.N., Park S.C., Woo J.R., Ma W., Liss M.A., Hirata T., Raheem O., Cacalano N.A., Kulidjian A.A, & Jamieson C.A. (2014). PCSD1, a new patient-derived model of bone metastatic prostate cancer, is castrate-resistant in the bone-niche. Journal of Translational Medicine, 12, 275.

Publication 2014

anti-PSA

Bath Cryosections Dry ice Edta Embedded paraffin Femoral tumors Formalin Frozen Isopentane Microscope Paraffin Spectrum analysis Tumors

Corresponding Organization :

Other organizations : University of California, San Diego, Wonkwang University, Okayama University, University of California, Los Angeles

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Variable analysis

independent variables

Intra-femoral tumors were fixed in 10% formalin, decalcified with 10% EDTA, embedded in OCT and frozen in isopentane/dry ice bath for cryosections or paraffin embedded
Subcutaneous tumors were fresh frozen in OCT or 10% formalin-fixed and embedded in paraffin at the time of harvest

dependent variables

IF and SC sections were H&E and immunostained with anti-PSA, 1:500 (DAKO)
Quantitation of PSA staining in FFPE sections was performed using The Spectrum Analysis algorithm package and ImageScope Analysis software

control variables

Five fields of view were randomly selected in Aperio ScanScope digital images at 20 X magnification in each of three different tumors in each treatment group

controls

Positive control: None mentioned
Negative control: None mentioned

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