Comprehensive Evaluation of PAXgene Fixation on Oral Cancer Tissues Using Routine Histology, Immunohistochemistry, and FTIR Microspectroscopy
The choice of tissue fixation is critical for preserving the morphology and biochemical information of tissues. Fragile oral tissues with lower tensile strength are challenging to process for histological applications as they are prone to processing damage, such as tissue tear, wrinkling, and tissue...
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| Format: | Article |
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MDPI AG
2021-06-01
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| Series: | Biomolecules |
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| Online Access: | https://www.mdpi.com/2218-273X/11/6/889 |
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| author | Pooja Lahiri Suranjana Mukherjee Biswajoy Ghosh Debnath Das Basudev Lahiri Shailendra Kumar Varshney Mousumi Pal Ranjan Rashmi Paul Jyotirmoy Chatterjee |
| author_facet | Pooja Lahiri Suranjana Mukherjee Biswajoy Ghosh Debnath Das Basudev Lahiri Shailendra Kumar Varshney Mousumi Pal Ranjan Rashmi Paul Jyotirmoy Chatterjee |
| author_sort | Pooja Lahiri |
| collection | DOAJ |
| description | The choice of tissue fixation is critical for preserving the morphology and biochemical information of tissues. Fragile oral tissues with lower tensile strength are challenging to process for histological applications as they are prone to processing damage, such as tissue tear, wrinkling, and tissue fall-off from slides. This leads to loss of morphological information and unnecessary delay in experimentation. In this study, we have characterized the new PAXgene tissue fixation system on oral buccal mucosal tissue of cancerous and normal pathology for routine histological and immunohistochemical applications. We aimed to minimize the processing damage of tissues and improve the quality of histological experiments. We also examined the preservation of biomolecules by PAXgene fixation using FTIR microspectroscopy. Our results demonstrate that the PAXgene-fixed tissues showed significantly less tissue fall-off from slides. Hematoxylin and Eosin staining showed comparable morphology between formalin-fixed and PAXgene-fixed tissues. Good quality and slightly superior immunostaining for cancer-associated proteins p53 and CK5/6 were observed in PAXgene-fixed tissues without antigen retrieval than formalin-fixed tissues. Further, FTIR measurements revealed superior preservation of glycogen, fatty acids, and amide III protein secondary structures in PAXgene-fixed tissues. Overall, we present the first comprehensive evaluation of the PAXgene tissue fixation system in oral tissues. This study concludes that the PAXgene tissue fixation system can be applied to oral tissues to perform diagnostic molecular pathology experiments without compromising the quality of the morphology or biochemistry of biomolecules. |
| first_indexed | 2024-03-10T10:23:39Z |
| format | Article |
| id | doaj.art-9ad7d697db1148be85d229ed55ec8273 |
| institution | Directory Open Access Journal |
| issn | 2218-273X |
| language | English |
| last_indexed | 2024-03-10T10:23:39Z |
| publishDate | 2021-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biomolecules |
| spelling | doaj.art-9ad7d697db1148be85d229ed55ec82732023-11-22T00:13:27ZengMDPI AGBiomolecules2218-273X2021-06-0111688910.3390/biom11060889Comprehensive Evaluation of PAXgene Fixation on Oral Cancer Tissues Using Routine Histology, Immunohistochemistry, and FTIR MicrospectroscopyPooja Lahiri0Suranjana Mukherjee1Biswajoy Ghosh2Debnath Das3Basudev Lahiri4Shailendra Kumar Varshney5Mousumi Pal6Ranjan Rashmi Paul7Jyotirmoy Chatterjee8School of Medical Science and Technology, Indian Institute of Technology Kharagpur, Kharagpur 721302, IndiaSchool of Medical Science and Technology, Indian Institute of Technology Kharagpur, Kharagpur 721302, IndiaSchool of Medical Science and Technology, Indian Institute of Technology Kharagpur, Kharagpur 721302, IndiaSchool of Medical Science and Technology, Indian Institute of Technology Kharagpur, Kharagpur 721302, IndiaDepartment of Electronics and Electrical Communication Engineering, Indian Institute of Technology Kharagpur, Kharagpur 721302, IndiaDepartment of Electronics and Electrical Communication Engineering, Indian Institute of Technology Kharagpur, Kharagpur 721302, IndiaGuru Nanak Institute of Dental Sciences and Research (GNIDSR), Kolkata 700114, IndiaDepartment of Oral & Dental Sciences, JIS University, Kolkata 700109, IndiaSchool of Medical Science and Technology, Indian Institute of Technology Kharagpur, Kharagpur 721302, IndiaThe choice of tissue fixation is critical for preserving the morphology and biochemical information of tissues. Fragile oral tissues with lower tensile strength are challenging to process for histological applications as they are prone to processing damage, such as tissue tear, wrinkling, and tissue fall-off from slides. This leads to loss of morphological information and unnecessary delay in experimentation. In this study, we have characterized the new PAXgene tissue fixation system on oral buccal mucosal tissue of cancerous and normal pathology for routine histological and immunohistochemical applications. We aimed to minimize the processing damage of tissues and improve the quality of histological experiments. We also examined the preservation of biomolecules by PAXgene fixation using FTIR microspectroscopy. Our results demonstrate that the PAXgene-fixed tissues showed significantly less tissue fall-off from slides. Hematoxylin and Eosin staining showed comparable morphology between formalin-fixed and PAXgene-fixed tissues. Good quality and slightly superior immunostaining for cancer-associated proteins p53 and CK5/6 were observed in PAXgene-fixed tissues without antigen retrieval than formalin-fixed tissues. Further, FTIR measurements revealed superior preservation of glycogen, fatty acids, and amide III protein secondary structures in PAXgene-fixed tissues. Overall, we present the first comprehensive evaluation of the PAXgene tissue fixation system in oral tissues. This study concludes that the PAXgene tissue fixation system can be applied to oral tissues to perform diagnostic molecular pathology experiments without compromising the quality of the morphology or biochemistry of biomolecules.https://www.mdpi.com/2218-273X/11/6/889PAXgene tissue fixationformalin fixationoral epithelial dysplasiaoral squamous cell carcinomaimmunohistochemistryspectral histology |
| spellingShingle | Pooja Lahiri Suranjana Mukherjee Biswajoy Ghosh Debnath Das Basudev Lahiri Shailendra Kumar Varshney Mousumi Pal Ranjan Rashmi Paul Jyotirmoy Chatterjee Comprehensive Evaluation of PAXgene Fixation on Oral Cancer Tissues Using Routine Histology, Immunohistochemistry, and FTIR Microspectroscopy Biomolecules PAXgene tissue fixation formalin fixation oral epithelial dysplasia oral squamous cell carcinoma immunohistochemistry spectral histology |
| title | Comprehensive Evaluation of PAXgene Fixation on Oral Cancer Tissues Using Routine Histology, Immunohistochemistry, and FTIR Microspectroscopy |
| title_full | Comprehensive Evaluation of PAXgene Fixation on Oral Cancer Tissues Using Routine Histology, Immunohistochemistry, and FTIR Microspectroscopy |
| title_fullStr | Comprehensive Evaluation of PAXgene Fixation on Oral Cancer Tissues Using Routine Histology, Immunohistochemistry, and FTIR Microspectroscopy |
| title_full_unstemmed | Comprehensive Evaluation of PAXgene Fixation on Oral Cancer Tissues Using Routine Histology, Immunohistochemistry, and FTIR Microspectroscopy |
| title_short | Comprehensive Evaluation of PAXgene Fixation on Oral Cancer Tissues Using Routine Histology, Immunohistochemistry, and FTIR Microspectroscopy |
| title_sort | comprehensive evaluation of paxgene fixation on oral cancer tissues using routine histology immunohistochemistry and ftir microspectroscopy |
| topic | PAXgene tissue fixation formalin fixation oral epithelial dysplasia oral squamous cell carcinoma immunohistochemistry spectral histology |
| url | https://www.mdpi.com/2218-273X/11/6/889 |
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