Skip to main content
NCBI home page
Molecular Pathology : MP logoLink to Molecular Pathology : MP
. 1998 Aug;51(4):233–236. doi: 10.1136/mp.51.4.233

A simple combined microdissection and aspiration device for the rapid procurement of single cells from clinical peripheral blood smears.

C P Beltinger 1, K M Debatin 1
PMCID: PMC395645  PMID: 9893754

Abstract

Molecular analysis of cells from cytology specimens can help to establish a diagnosis in ambiguous cases. However, mutations in heterogeneous samples might not be detected because of the diluting effect of DNA from normal background cells. Even if a mutation were detected, it could not be traced back to a specific cell type. Molecular analysis of single cells circumvents this problem. Both mechanical and laser assisted methods have been described for the selective procurement of cells from histology slides; however, they have the drawback of either being technically demanding or expensive. Furthermore, it is nuclear whether they can be applied to cytology specimens. Finally, few of these techniques are able to procure single cells. Therefore, we developed a simplified combined microdissection and aspiration device for the rapid procurement of single cells from clinical cytology specimens. The principle of this device, called the cytopicker, is the combination of the microdissection tool, a steel cannula, with the aspiration tool, a glass capillary connected to a vacuum, into one device. Steel cannulae are optimal for microdissection of cells from the hard matrix of cytology specimens but aspirate poorly. On the other hand, glass capillaries are suboptimal for dissecting but aspirate very well. Combining both tools into one by inserting the capillary into the cannula allows optimal dissection using the cannula (with the glass capillary with-drawn and thus protected), followed by optimal aspiration using the capillary (after being advanced through the cannula). All movements of the device are controlled by just one micromanipulator, making the cytopicker inexpensive to manufacture. The cytopicker can rapidly and simply procure single cells, such as lymphoblasts, from cytology specimens, such as peripheral blood smears. DNA from these cells can be amplified by PCR. However, precautions have to be taken to avoid contamination. Once improved further, the cytopicker might facilitate molecular analysis in the routine cytology laboratory.

Full Text

The Full Text of this article is available as a PDF (171.3 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aguilar Martinez P., Jeanjean P., Masmejean C., Guillard A., Biron C., Rabesandratana H., Schved J. F. Simple and rapid detection of the newly described mutations in the HLA-H gene. Blood. 1997 Mar 1;89(5):1835–1836. [PubMed] [Google Scholar]
  2. Becker I., Becker K. F., Röhrl M. H., Minkus G., Schütze K., Höfler H. Single-cell mutation analysis of tumors from stained histologic slides. Lab Invest. 1996 Dec;75(6):801–807. [PubMed] [Google Scholar]
  3. Beltinger C. P., Klimek F., Debatin K. M. Whole genome amplification of single cells from clinical peripheral blood smears. Mol Pathol. 1997 Oct;50(5):272–275. doi: 10.1136/mp.50.5.272. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bennett J. M., Catovsky D., Daniel M. T., Flandrin G., Galton D. A., Gralnick H. R., Sultan C. The morphological classification of acute lymphoblastic leukaemia: concordance among observers and clinical correlations. Br J Haematol. 1981 Apr;47(4):553–561. doi: 10.1111/j.1365-2141.1981.tb02684.x. [DOI] [PubMed] [Google Scholar]
  5. Beutler E., Gelbart T., West C., Lee P., Adams M., Blackstone R., Pockros P., Kosty M., Venditti C. P., Phatak P. D. Mutation analysis in hereditary hemochromatosis. Blood Cells Mol Dis. 1996;22(2):187–194b. doi: 10.1006/bcmd.1996.0027. [DOI] [PubMed] [Google Scholar]
  6. Beutler E. Genetic irony beyond haemochromatosis: clinical effects of HLA-H mutations. Lancet. 1997 Feb 1;349(9048):296–297. doi: 10.1016/S0140-6736(97)22005-2. [DOI] [PubMed] [Google Scholar]
  7. Böhm M., Wieland I., Schütze K., Rübben H. Microbeam MOMeNT: non-contact laser microdissection of membrane-mounted native tissue. Am J Pathol. 1997 Jul;151(1):63–67. [PMC free article] [PubMed] [Google Scholar]
  8. Emmert-Buck M. R., Bonner R. F., Smith P. D., Chuaqui R. F., Zhuang Z., Goldstein S. R., Weiss R. A., Liotta L. A. Laser capture microdissection. Science. 1996 Nov 8;274(5289):998–1001. doi: 10.1126/science.274.5289.998. [DOI] [PubMed] [Google Scholar]
  9. Feder J. N., Gnirke A., Thomas W., Tsuchihashi Z., Ruddy D. A., Basava A., Dormishian F., Domingo R., Jr, Ellis M. C., Fullan A. A novel MHC class I-like gene is mutated in patients with hereditary haemochromatosis. Nat Genet. 1996 Aug;13(4):399–408. doi: 10.1038/ng0896-399. [DOI] [PubMed] [Google Scholar]
  10. Going J. J., Lamb R. F. Practical histological microdissection for PCR analysis. J Pathol. 1996 May;179(1):121–124. doi: 10.1002/(SICI)1096-9896(199605)179:1<121::AID-PATH536>3.0.CO;2-D. [DOI] [PubMed] [Google Scholar]
  11. Guttridge M. G., Burr C., Klouda P. T. Identification of HLA-B35, B53, B18, B5, B78, and B17 alleles by the polymerase chain reaction using sequence-specific primers (PCR-SSP). Tissue Antigens. 1994 Jul;44(1):43–46. doi: 10.1111/j.1399-0039.1994.tb02355.x. [DOI] [PubMed] [Google Scholar]
  12. Jazwinska E. C., Cullen L. M., Busfield F., Pyper W. R., Webb S. I., Powell L. W., Morris C. P., Walsh T. P. Haemochromatosis and HLA-H. Nat Genet. 1996 Nov;14(3):249–251. doi: 10.1038/ng1196-249. [DOI] [PubMed] [Google Scholar]
  13. Jouanolle A. M., Gandon G., Jézéquel P., Blayau M., Campion M. L., Yaouanq J., Mosser J., Fergelot P., Chauvel B., Bouric P. Haemochromatosis and HLA-H. Nat Genet. 1996 Nov;14(3):251–252. doi: 10.1038/ng1196-251. [DOI] [PubMed] [Google Scholar]
  14. Merryweather-Clarke A. T., Pointon J. J., Shearman J. D., Robson K. J. Global prevalence of putative haemochromatosis mutations. J Med Genet. 1997 Apr;34(4):275–278. doi: 10.1136/jmg.34.4.275. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Miller S. A., Dykes D. D., Polesky H. F. A simple salting out procedure for extracting DNA from human nucleated cells. Nucleic Acids Res. 1988 Feb 11;16(3):1215–1215. doi: 10.1093/nar/16.3.1215. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Moskaluk C. A., Kern S. E. Microdissection and polymerase chain reaction amplification of genomic DNA from histological tissue sections. Am J Pathol. 1997 May;150(5):1547–1552. [PMC free article] [PubMed] [Google Scholar]
  17. Smillie D. A PCR-SSP method for detecting the Cys282Tyr mutation in the HFE gene associated with hereditary haemochromatosis. Mol Pathol. 1997 Oct;50(5):275–276. doi: 10.1136/mp.50.5.275. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Takeuchi T., Soejima H., Faed J. M., Yun K. Efficient large-scale screening for the hemochromatosis susceptibility gene mutation. Blood. 1997 Oct 1;90(7):2848–2849. [PubMed] [Google Scholar]
  19. Zhuang Z., Bertheau P., Emmert-Buck M. R., Liotta L. A., Gnarra J., Linehan W. M., Lubensky I. A. A microdissection technique for archival DNA analysis of specific cell populations in lesions < 1 mm in size. Am J Pathol. 1995 Mar;146(3):620–625. [PMC free article] [PubMed] [Google Scholar]
  20. Zhuang Z., Roth M. J., Emmert-Buck M. R., Lubensky I. A., Liotta L. A., Solomon D. Detection of the von Hippel-Lindau gene deletion in cytologic specimens using microdissection and the polymerase chain reaction. Acta Cytol. 1994 Sep-Oct;38(5):671–675. [PubMed] [Google Scholar]

Articles from Molecular Pathology are provided here courtesy of BMJ Publishing Group

RESOURCES