A Comparison of Different Sample Processing Protocols for MALDI Imaging Mass Spectrometry Analysis of Formalin-Fixed Multiple Myeloma Cells
Sample processing of formalin-fixed specimens constitutes a major challenge in molecular profiling efforts. Pre-analytical factors such as fixative temperature, dehydration, and embedding media affect downstream analysis, generating data dependent on technical processing rather than disease state. I...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-02-01
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| Series: | Cancers |
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| Online Access: | https://www.mdpi.com/2072-6694/15/3/974 |
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| author | Rita Casadonte Jörg Kriegsmann Mark Kriegsmann Katharina Kriegsmann Roberta Torcasio Maria Eugenia Gallo Cantafio Giuseppe Viglietto Nicola Amodio |
| author_facet | Rita Casadonte Jörg Kriegsmann Mark Kriegsmann Katharina Kriegsmann Roberta Torcasio Maria Eugenia Gallo Cantafio Giuseppe Viglietto Nicola Amodio |
| author_sort | Rita Casadonte |
| collection | DOAJ |
| description | Sample processing of formalin-fixed specimens constitutes a major challenge in molecular profiling efforts. Pre-analytical factors such as fixative temperature, dehydration, and embedding media affect downstream analysis, generating data dependent on technical processing rather than disease state. In this study, we investigated two different sample processing methods, including the use of the cytospin sample preparation and automated sample processing apparatuses for proteomic analysis of multiple myeloma (MM) cell lines using imaging mass spectrometry (IMS). In addition, two sample-embedding instruments using different reagents and processing times were considered. Three MM cell lines fixed in 4% paraformaldehyde were either directly centrifuged onto glass slides using cytospin preparation techniques or processed to create paraffin-embedded specimens with an automatic tissue processor, and further cut onto glass slides for IMS analysis. The number of peaks obtained from paraffin-embedded samples was comparable between the two different sample processing instruments. Interestingly, spectra profiles showed enhanced ion yield in cytospin compared to paraffin-embedded samples along with high reproducibility compared to the sample replicate. |
| first_indexed | 2024-03-11T09:49:47Z |
| format | Article |
| id | doaj.art-178a45b8d660469a8307af4c58d1be57 |
| institution | Directory Open Access Journal |
| issn | 2072-6694 |
| language | English |
| last_indexed | 2024-03-11T09:49:47Z |
| publishDate | 2023-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cancers |
| spelling | doaj.art-178a45b8d660469a8307af4c58d1be572023-11-16T16:19:54ZengMDPI AGCancers2072-66942023-02-0115397410.3390/cancers15030974A Comparison of Different Sample Processing Protocols for MALDI Imaging Mass Spectrometry Analysis of Formalin-Fixed Multiple Myeloma CellsRita Casadonte0Jörg Kriegsmann1Mark Kriegsmann2Katharina Kriegsmann3Roberta Torcasio4Maria Eugenia Gallo Cantafio5Giuseppe Viglietto6Nicola Amodio7Proteopath GmbH, 54296 Trier, GermanyProteopath GmbH, 54296 Trier, GermanyInstitute of Pathology, University Hospital Heidelberg, 69120 Heidelberg, GermanyDepartment of Hematology, Oncology and Rheumatology, Heidelberg University, 69120 Heidelberg, GermanyDepartment of Experimental and Clinical Medicine, University Magna Graecia of Catanzaro, 88100 Catanzaro, ItalyDepartment of Experimental and Clinical Medicine, University Magna Graecia of Catanzaro, 88100 Catanzaro, ItalyDepartment of Experimental and Clinical Medicine, University Magna Graecia of Catanzaro, 88100 Catanzaro, ItalyDepartment of Experimental and Clinical Medicine, University Magna Graecia of Catanzaro, 88100 Catanzaro, ItalySample processing of formalin-fixed specimens constitutes a major challenge in molecular profiling efforts. Pre-analytical factors such as fixative temperature, dehydration, and embedding media affect downstream analysis, generating data dependent on technical processing rather than disease state. In this study, we investigated two different sample processing methods, including the use of the cytospin sample preparation and automated sample processing apparatuses for proteomic analysis of multiple myeloma (MM) cell lines using imaging mass spectrometry (IMS). In addition, two sample-embedding instruments using different reagents and processing times were considered. Three MM cell lines fixed in 4% paraformaldehyde were either directly centrifuged onto glass slides using cytospin preparation techniques or processed to create paraffin-embedded specimens with an automatic tissue processor, and further cut onto glass slides for IMS analysis. The number of peaks obtained from paraffin-embedded samples was comparable between the two different sample processing instruments. Interestingly, spectra profiles showed enhanced ion yield in cytospin compared to paraffin-embedded samples along with high reproducibility compared to the sample replicate.https://www.mdpi.com/2072-6694/15/3/974cytospinFFPEimaging mass spectrometryMALDIsample preparation |
| spellingShingle | Rita Casadonte Jörg Kriegsmann Mark Kriegsmann Katharina Kriegsmann Roberta Torcasio Maria Eugenia Gallo Cantafio Giuseppe Viglietto Nicola Amodio A Comparison of Different Sample Processing Protocols for MALDI Imaging Mass Spectrometry Analysis of Formalin-Fixed Multiple Myeloma Cells Cancers cytospin FFPE imaging mass spectrometry MALDI sample preparation |
| title | A Comparison of Different Sample Processing Protocols for MALDI Imaging Mass Spectrometry Analysis of Formalin-Fixed Multiple Myeloma Cells |
| title_full | A Comparison of Different Sample Processing Protocols for MALDI Imaging Mass Spectrometry Analysis of Formalin-Fixed Multiple Myeloma Cells |
| title_fullStr | A Comparison of Different Sample Processing Protocols for MALDI Imaging Mass Spectrometry Analysis of Formalin-Fixed Multiple Myeloma Cells |
| title_full_unstemmed | A Comparison of Different Sample Processing Protocols for MALDI Imaging Mass Spectrometry Analysis of Formalin-Fixed Multiple Myeloma Cells |
| title_short | A Comparison of Different Sample Processing Protocols for MALDI Imaging Mass Spectrometry Analysis of Formalin-Fixed Multiple Myeloma Cells |
| title_sort | comparison of different sample processing protocols for maldi imaging mass spectrometry analysis of formalin fixed multiple myeloma cells |
| topic | cytospin FFPE imaging mass spectrometry MALDI sample preparation |
| url | https://www.mdpi.com/2072-6694/15/3/974 |
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