Use of bovine serum albumin might impair immunofluorescence signal in thick tissue samples
Abstract Significant progress in microscopic imaging techniques allowed transition from predominantly qualitative methods to a powerful tool for quantitative research, driven by improved instrumentation and computational power. Furthermore, previously limited to thin, laser-permeable tissue sections...
Saved in:
| Main Authors: | , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Nature Portfolio
2025-07-01
|
| Series: | Scientific Reports |
| Subjects: | |
| Online Access: | https://doi.org/10.1038/s41598-025-06876-z |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1849402928623779840 |
|---|---|
| author | Anna Chwastowicz Artur Wolny Magdalena Sobień Marcin Barański Jacek Tomczuk Michał Szatkowski Aleksandra Szredzka Jakub Gołąb Leszek Kaczmarek Marzena Stefaniuk Paweł Matryba |
| author_facet | Anna Chwastowicz Artur Wolny Magdalena Sobień Marcin Barański Jacek Tomczuk Michał Szatkowski Aleksandra Szredzka Jakub Gołąb Leszek Kaczmarek Marzena Stefaniuk Paweł Matryba |
| author_sort | Anna Chwastowicz |
| collection | DOAJ |
| description | Abstract Significant progress in microscopic imaging techniques allowed transition from predominantly qualitative methods to a powerful tool for quantitative research, driven by improved instrumentation and computational power. Furthermore, previously limited to thin, laser-permeable tissue sections, imaging techniques have been revolutionized by the advent of tissue optical clearing. This innovation enables the visualization and quantitative analysis of entire organs and even whole bodies at cellular resolution. However, achieving high-quality imaging depends not only on the transparency of the tissue preparation but also on precise immunofluorescence labeling to ensure accurate signal detection and reliable study outcomes. In this study, we evaluated whether various reagents that are typically applied during the tissue blocking step prior to immunofluorescence staining affect the quality of the obtained image in thick and optically cleared samples. We demonstrate that the commonly employed tissue blocking step does not improve imaging conditions and even can substantially degrade fluorescence signal quality, particularly in large, optically cleared tissues such as whole mouse brain hemispheres. |
| format | Article |
| id | doaj-art-d2a748293fec4d69b59657d8cdca0d00 |
| institution | Kabale University |
| issn | 2045-2322 |
| language | English |
| publishDate | 2025-07-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj-art-d2a748293fec4d69b59657d8cdca0d002025-08-20T03:37:24ZengNature PortfolioScientific Reports2045-23222025-07-0115111110.1038/s41598-025-06876-zUse of bovine serum albumin might impair immunofluorescence signal in thick tissue samplesAnna Chwastowicz0Artur Wolny1Magdalena Sobień2Marcin Barański3Jacek Tomczuk4Michał Szatkowski5Aleksandra Szredzka6Jakub Gołąb7Leszek Kaczmarek8Marzena Stefaniuk9Paweł Matryba10Department of Immunology, Medical University of WarsawLaboratory of Imaging Tissue Structure and Function, Nencki Institute of Experimental Biology of Polish Academy of SciencesLaboratory of Neurobiology, BRAINCITY, Nencki Institute of Experimental Biology of Polish Academy of SciencesLaboratory of Neurobiology, BRAINCITY, Nencki Institute of Experimental Biology of Polish Academy of SciencesDepartment of Immunology, Medical University of WarsawDepartment of Immunology, Medical University of WarsawDepartment of Immunology, Medical University of WarsawDepartment of Immunology, Medical University of WarsawLaboratory of Neurobiology, BRAINCITY, Nencki Institute of Experimental Biology of Polish Academy of SciencesLaboratory of Neurobiology, BRAINCITY, Nencki Institute of Experimental Biology of Polish Academy of SciencesDepartment of Immunology, Medical University of WarsawAbstract Significant progress in microscopic imaging techniques allowed transition from predominantly qualitative methods to a powerful tool for quantitative research, driven by improved instrumentation and computational power. Furthermore, previously limited to thin, laser-permeable tissue sections, imaging techniques have been revolutionized by the advent of tissue optical clearing. This innovation enables the visualization and quantitative analysis of entire organs and even whole bodies at cellular resolution. However, achieving high-quality imaging depends not only on the transparency of the tissue preparation but also on precise immunofluorescence labeling to ensure accurate signal detection and reliable study outcomes. In this study, we evaluated whether various reagents that are typically applied during the tissue blocking step prior to immunofluorescence staining affect the quality of the obtained image in thick and optically cleared samples. We demonstrate that the commonly employed tissue blocking step does not improve imaging conditions and even can substantially degrade fluorescence signal quality, particularly in large, optically cleared tissues such as whole mouse brain hemispheres.https://doi.org/10.1038/s41598-025-06876-zTissue clearingConfocal microscopyBSABovine serum albuminImmunofluorescence |
| spellingShingle | Anna Chwastowicz Artur Wolny Magdalena Sobień Marcin Barański Jacek Tomczuk Michał Szatkowski Aleksandra Szredzka Jakub Gołąb Leszek Kaczmarek Marzena Stefaniuk Paweł Matryba Use of bovine serum albumin might impair immunofluorescence signal in thick tissue samples Scientific Reports Tissue clearing Confocal microscopy BSA Bovine serum albumin Immunofluorescence |
| title | Use of bovine serum albumin might impair immunofluorescence signal in thick tissue samples |
| title_full | Use of bovine serum albumin might impair immunofluorescence signal in thick tissue samples |
| title_fullStr | Use of bovine serum albumin might impair immunofluorescence signal in thick tissue samples |
| title_full_unstemmed | Use of bovine serum albumin might impair immunofluorescence signal in thick tissue samples |
| title_short | Use of bovine serum albumin might impair immunofluorescence signal in thick tissue samples |
| title_sort | use of bovine serum albumin might impair immunofluorescence signal in thick tissue samples |
| topic | Tissue clearing Confocal microscopy BSA Bovine serum albumin Immunofluorescence |
| url | https://doi.org/10.1038/s41598-025-06876-z |
| work_keys_str_mv | AT annachwastowicz useofbovineserumalbuminmightimpairimmunofluorescencesignalinthicktissuesamples AT arturwolny useofbovineserumalbuminmightimpairimmunofluorescencesignalinthicktissuesamples AT magdalenasobien useofbovineserumalbuminmightimpairimmunofluorescencesignalinthicktissuesamples AT marcinbaranski useofbovineserumalbuminmightimpairimmunofluorescencesignalinthicktissuesamples AT jacektomczuk useofbovineserumalbuminmightimpairimmunofluorescencesignalinthicktissuesamples AT michałszatkowski useofbovineserumalbuminmightimpairimmunofluorescencesignalinthicktissuesamples AT aleksandraszredzka useofbovineserumalbuminmightimpairimmunofluorescencesignalinthicktissuesamples AT jakubgołab useofbovineserumalbuminmightimpairimmunofluorescencesignalinthicktissuesamples AT leszekkaczmarek useofbovineserumalbuminmightimpairimmunofluorescencesignalinthicktissuesamples AT marzenastefaniuk useofbovineserumalbuminmightimpairimmunofluorescencesignalinthicktissuesamples AT pawełmatryba useofbovineserumalbuminmightimpairimmunofluorescencesignalinthicktissuesamples |