Immunohistochemical Analysis of FHL2 in Ovarian Tissues

The expression of FHL2 protein in paraffin-embedded human ovarian tissues and GCTs was detected by a previously described peroxidase-based immunohistochemistry protocol.^{35 (link)} Immunosignals were visualized with a DAB kit (Invitrogen, Grand Island, NY, USA). The sections were counterstained with Mayer's hematoxylin and scanned with an iSCAN Coreo Slide Scaner (Ventana Medical Systems, Inc., Oro Valley, AZ, USA). The positivity (i.e., the number of positively stained cells relative to the total number of cells in the tissue section) and the intensity of the positive immunosignals were quantified with Aperio ImageScope software (Leica Biosystems Inc., Buffalo Grove, IL, USA).

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Hua G., He C., Lv X., Fan L., Wang C., Remmenga S.W., Rodabaugh K.J., Yang L., Lele S.M., Yang P., Karpf A.R., Davis J.S, & Wang C. (2016). The four and a half LIM domains 2 (FHL2) regulates ovarian granulosa cell tumor progression via controlling AKT1 transcription. Cell Death & Disease, 7(7), e2297-.

Publication 2016

DAB kit iSCAN Coreo Slide Scaner Aperio ImageScope software

Buffalo Fhl2 Hematoxylin Human Immunohistochemistry Iscan Ovarian Paraffin Peroxidase Protein Tissue

Corresponding Organization :

Other organizations : University of Nebraska Medical Center, Huazhong Agricultural University, University of Maryland, Baltimore

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Expression of FHL2 protein
Positivity (number of positively stained cells relative to total cells)
Intensity of positive immunosignals

control variables

Paraffin-embedded human ovarian tissues and GCTs (Granulosa Cell Tumors)
Peroxidase-based immunohistochemistry protocol
DAB kit (Invitrogen, Grand Island, NY, USA) for visualizing immunosignals
Counterstaining with Mayer's hematoxylin
Scanning with iSCAN Coreo Slide Scaner (Ventana Medical Systems, Inc., Oro Valley, AZ, USA)
Quantification with Aperio ImageScope software (Leica Biosystems Inc., Buffalo Grove, IL, USA)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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