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| Content Provider | PubMed Central |
|---|---|
| Author | Woltmann, G. Ward, R. Symon, F. Rew, D. Wardlaw, A. Pavord, I. |
| Abstract | BACKGROUND—Sputuminduction is an important non-invasive technique for measuring airwayinflammation in asthma. Cell numbers are often too low for flowcytometric analysis. Laser scanning cytometry (LSC) is a noveltechnique that allows objective multicolour fluorescence analysis ofcells on a microscope slide. METHODS—LSC was usedto determine sputum eosinophil and bronchial epithelial cell counts. Wefirst confirmed that we could measure eosinophil counts accurately inperipheral blood using α-major basic protein (MBP) immunofluorescentstaining. Sputum induction was performed according to standardprotocols. Sputum samples from eight normal controls and 12 asthmaticpatients were analysed by LSC and manual counting by two independentobservers. Octospot cytospins were fixed and stained withmouse-α-human-MBP monoclonal antibody or mouse-α-human-cytokeratinantibody and goat-α-mouse Oregon Green conjugated second antibody. RESULTS—Sputuminduction provided a mean (SE) of 0.99 (0.2) × 106 cellsper donor. More than 3000 cells on three cytospins per slide wereanalysed per cell type. The intraclass correlation coefficient (R) andstandard deviation (SD) of differences in eosinophils determined bymanual counting and LSC were 0.9 and 2.1, respectively, and forbronchial epithelial cell counts they were 0.7 and 2.0.Selectivedetection of labelled cells was confirmed visually after relocation. CONCLUSION—Eosinophilsand bronchial epithelial cells can be accurately and reproduciblycounted in an objective manner. LSC is therefore a potentially powerfulnew method for immunophenotyping leucocytes and epithelial cellsobjectively in induced sputum in patients with asthma. |
| Starting Page | 124 |
| File Format | |
| ISSN | 14683296 |
| e-ISSN | 14683296 |
| Journal | Thorax |
| Issue Number | 2 |
| Volume Number | 54 |
| Language | English |
| Publisher | BMJ Group |
| Publisher Date | 1999-02-01 |
| Access Restriction | Open |
| Rights Holder | BMJ Group |
| Subject Keyword | Research in Higher Education |
| Content Type | Text |
| Resource Type | Article |
| Subject | Pulmonary and Respiratory Medicine |
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