A study of detection and comparison of immunofluorescence on formalin-fixed paraffin-embedded tissue with fresh frozen renal biopsy specimen

Solanki, Ranjana; Solanki, Madan Kumar; Hemrajani, Deepika; Saha, Jayesh

doi:10.4103/1319-2442.256842

Cited by 5 publications

(7 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Several techniques, including enzyme digestion and heat treatment, have been tried to reveal the antigen. For antigen retrieval, the enzymes trypsin, pepsin, protease VII, pronase, protease XXIV, and proteinase K have been utilized for varying amounts of time, temperature, and concentration [ 5 ].…”

Section: Discussionmentioning

confidence: 99%

“…In a study by Solanki et al, 92% of cases were diagnosed based on immunofluorescence done on formalin-fixed, paraffin-embedded tissue by proteinase K antigen retrieval on kidney biopsies compared to immunofluorescence done on frozen sections [ 5 ]. A similar methodology was also used by Wagrowska-Danilewicz et al, Nada et al, Singh et al, and Mohammadzadeh et al [ 2 , 6 - 8 ].…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

A Comparative Study of Immunofluorescence on Formalin-Fixed, Paraffin-Embedded Versus Fresh Frozen Kidney Biopsy

Das¹,

Lakadong²,

Dey³

et al. 2023

Cureus

View full text Add to dashboard Cite

Background Immunofluorescence techniques done on formalin-fixed, paraffin-embedded tissue can serve as salvage techniques in cases where immunofluorescence on the frozen section may not be adequate or available. The present study was undertaken to assess the diagnostic utility of paraffin immunofluorescence by proteinase K digestion on renal biopsy compared to fresh frozen immunofluorescence. Methodology The paraffin immunofluorescence by proteinase K digestion of paraffin-embedded renal biopsy (IF-FFPE) was standardized and compared with the immunofluorescence on fresh frozen tissue (IF-Frozen). A total of 50 cases of the native renal biopsy were included in the study, and their intensity for fluorescein isothiocyanate-labeled IgA, IgG, IgM, C3, kappa, and lambda was compared. Results A total of 50 cases of the native renal biopsy were included in the study, and their intensity for fluorescein isothiocyanate-labeled antibodies of IgA, IgG, IgM, C3, kappa, and lambda was compared. The difference of 2+ intensity of antibodies between IF-FFPE and IF-Frozen was noted mainly in lupus nephritis (15%), followed by IgA nephropathy (10%) and membranoproliferative glomerulonephritis (7%). IF-FFPE showed a sensitivity of 90.3%, 91.8%, 82.7%, 81.1%, 92.1%, and 94.6% for IgA, IgG, IgM, C3, kappa, and lambda, respectively, whereas specificity was 100% for IgA, IgG, C3, kappa, and lambda and 95.2% for IgM. Conclusions Immunofluorescence techniques done on formalin-fixed, paraffin-embedded tissue can serve as salvage techniques in kidney biopsies.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

A Comparative Study of Immunofluorescence on Formalin-Fixed, Paraffin-Embedded Versus Fresh Frozen Kidney Biopsy

Das¹,

Lakadong²,

Dey³

et al. 2023

Cureus

View full text Add to dashboard Cite

show abstract

“…1F). Owing to the fluorescence quenching of GFP by the high temperatures used in antigen retrieval,11,22 we opted for other methods with proven value for immunofluorescence, such as pepsase and tryptase 9,23. Enzymatic digestion helps to break down the protein cross-links formed during formalin or paraformaldehyde fixation and exposes the antigenic immune complexes to staining.…”

Section: Discussionmentioning

confidence: 99%

“…Owing to the fluorescence quenching of GFP by the high temperatures used in antigen retrieval, 11,22 we opted for other methods with proven value for immunofluorescence, such as pepsase and tryptase. 9,23 Enzymatic digestion helps to break down the protein cross-links formed during formalin or paraformaldehyde fixation and exposes the antigenic immune complexes to staining. However, in our study, these methods did not work very well to expose the antibody binding sites of GFAP and Iba1, especially with tryptase, and the results were similar to those of negative controls.…”

Section: Discussionmentioning

confidence: 99%

Detecting Green Fluorescent Protein-tagged Cryptococcus neoformans by Immunofluorescence on Paraffin-embedded Brain Sections

Chen

Yingcai

Zhu

2021

Applied Immunohistochemistry &Amp; Molecular Morphology

View full text Add to dashboard Cite

Cryptococcus neoformans is an important pathogen causing opportunistic fungal meningitis. The pathogenic mechanism of cryptococcal meningitis remains unclear. We aimed to describe a practical approach for studying the pathologic features of cryptococcal central nervous system infection by immunofluorescence on paraffin-embedded brain of mice using different antigen retrieval methods. After 14 days of intratracheal inoculation of green fluorescent protein-tagged C. neoformans (H99-GFP), C57BL/6J mice brains were fixed in 4% paraformaldehyde and embedded in paraffin. Antigen retrieval methods such as microwaves, 1% sodium lauryl sulfate, 1 N HCl, pepsase, and tryptase were used on 5-μm paraffin sections and the effects were compared. The green fluorescence of H99-GFP persisted with antigen retrieval using 1% sodium lauryl sulfate. After immunofluorescent staining, H99-GFP, glial fibrillary acidic protein-tagged astrocytes, and ionized calcium-binding adapter molecule 1-tagged microglia could be observed clearly. Based on our results, we provide a practical approach for the further study of the interaction between C. neoformans and brain cells.

show abstract

“…The main disadvantage of formaldehyde fixation and paraffin embedding is the denaturation of tissue antigens, leading to the increased difficulty of IF antigen investigation [ 10 ]. Apart from its major role in renal pathology as a salvage method, recent evidence indicates that IF-P is almost sensitive and specific in cases where cortical tissue for immunofluorescence investigations is unavailable due to technical factors; however, it may be more difficult to detect C3 [ 11 , 12 ].…”

Section: Introductionmentioning

confidence: 99%

Diagnostic Accuracy of Direct Immunofluorescence Test on Paraffin-Embedded Blocks in Comparison with Frozen Section Blocks in Renal Biopsies

Mohammadzadeh

Aghakhaninejad

Azad

et al. 2022

International Journal of Nephrology

View full text Add to dashboard Cite

Background. In several published research, the evaluation of renal disorders using immunofluorescence on formalin-fixed, paraffin-embedded (FFPE) tissue sections versus immunofluorescence on frozen sections was compared. Each technique’s accuracy varies greatly. This study’s objective was to assess IF-P as a potential replacement for IF-F in the diagnosis of renal biopsy specimens. Materials and Methods. To show immunoglobulin IgA, IgG, IgM, and C3 immune deposits, proteinase K digestion of paraffin-embedded renal biopsy was standardized and used in 51 renal biopsies. Sensitivity, specificity, false-positive, and false-negative values were calculated. Results. IF-P showed a sensitivity of 93.1%, 76.9%, 63.6%, and 33.3%, and a specificity of 100%, 97.3%, 95%, and 100% for IgG, IgA, IgM, and C3, respectively. Compared to cases that had both routine IF and IF-P, 50 of 51 showed either the same amount of staining for the diagnostic immunoglobulin/complement or a small amount of difference. In most of the cases (49 of 51), diagnostic findings were found. Conclusion. IF-P is a sensitive and precise approach for assessing immune deposits in renal tissue biopsies. We come to the conclusion that IF-P serves as a beneficial salvage immunohistochemistry method for renal biopsies that do not contain enough cortical tissue for IF-F.

show abstract

A study of detection and comparison of immunofluorescence on formalin-fixed paraffin-embedded tissue with fresh frozen renal biopsy specimen

Cited by 5 publications

References 0 publications

A Comparative Study of Immunofluorescence on Formalin-Fixed, Paraffin-Embedded Versus Fresh Frozen Kidney Biopsy

A Comparative Study of Immunofluorescence on Formalin-Fixed, Paraffin-Embedded Versus Fresh Frozen Kidney Biopsy

Detecting Green Fluorescent Protein-tagged Cryptococcus neoformans by Immunofluorescence on Paraffin-embedded Brain Sections

Diagnostic Accuracy of Direct Immunofluorescence Test on Paraffin-Embedded Blocks in Comparison with Frozen Section Blocks in Renal Biopsies

Contact Info

Product

Resources

About