The cytologic diagnosis of Mycobacterium kansasi tuberculosis by fluorescence microscopy of Papanicolaou-stained specimens

The cytologic diagnosis of Mycobacterium kansasi tuberculosis by fluorescence microscopy of Papanicolaou-stained specimens
The sensitivities of (i) Papanicolaou fluorescence, (ii) auramine rhodamine fluorescence, and (iii) Ziehl-Neelsen staining were compared for their ability to detect the atypical mycobacterium Myco. kansasi in cytological samples. Ninety-two cases were investigated, and the sensitivities of the three methods of detection were found to be 36.9%, 12.0%, and 20.7%, respectively. The control groups consisted of 30 specimens from cases of bronchial carcinoma and 30 of pneumonia. All cases were proved by microbiology. No false-positive results were recorded using Papanicolaou fluorescence. An important but coincidental finding arising from this study was that infection by the atypical mycobacterium Myco. kansasi causes cytological patterns corresponding to those normally associated with acute pneumonia and not to tuberculosis.     

Detection of Charcot-Leyden crystals by fluorescence microscopy of Papanicolaou-stained smears of sputum, bronchoalveolar lavage fluid, and bronchial secretions

Fluorescence microscopy was used to examine Papanicolaou-stained smears of sputum and other secretions from the respiratory tract. Under these conditions Charcot-Leyden crystals (CLC) appear as bright yellow-green fluorescing needles. the study was performed to determine the value of this approach for the diagnosis of allergic lung diseases. the time taken to detect the crystals was recorded and the sensitivity of fluorescence microscopy for the detection of CLC was compared with light microscopy of the same samples. the data show that fluorescence microscopy is superior to light microscopy for the detection of CLC. the characteristic needle-shaped crystal can be recognized easily and fragments of crystals could be easily identified. In doubtful cases of allergic lung diseases, fluorescence microscopy may be used to supplement light microscopy for the detection of Charcot-Leyden crystals.     

Aspergillus in the Papanicolaou stain: morphology, fluorescence and diagnostic feasibility

hettlich c., küpper th., wehle k. and pfitzer p. (1998) Cytopathology 9, 381–388
Aspergillus in the Papanicolaou stain: morphology, fluorescence and diagnostic feasibility
Aspergillus species exhibit a distinct and clear fluorescence in Papanicolaou-stained cytological samples. The Papanicolaou (PAP) stain enhances the autofluorescence of cultured aspergilli and allows better cytological recognition of the fungus by fluorescence microscopy when it is not easily discerned from its surroundings by light microscopy. Morphological properties can be better distinguished and facilitate the differentiation of aspergillus organisms from other filamentous fungi. Neither light nor fluorescence microscopy, the cytological quality nor the presence of phagocytosed hyphae in alveolar macrophages allow distinction between infection and contamination with Aspergillus species. Only the presence of eosinophilic inflammation permits a tentative diagnosis of an Aspergillus infection. In conclusion, PAP fluorescence reduces the need for special stains, is superior to and quicker than other investigative techniques and enhances the sensitivity and specificity of cytological investigation when a rapid and reliable identification of Aspergillus is needed.     

The Cytological Diagnosis of Pneumocystis Carinii By Fluorescence Microscopy of Papanicolaou Stained Bronchoalveolar Lavage Specimens

In a retrospective and prospective analysis fluorescence microscopy of Papanicolaou stained bronchoalveolar lavage specimens has been applied to the diagnosis of Pneumocystis carinii (PC) in routine cytology. The pneumocysts presented as circular structures of 5 microns in diameter and of brilliant green-yellow fluorescence surrounding two mirror image reniform structures. Fluorescent inclusions of 1-3 microns diameter within the alveolar macrophages could be identified as remnants of pneumocysts by a follow-up of all steps of degradation ending in very small irregular granules. By applying both criteria, i.e. pneumocysts with reniform bodies and degradation inclusions within macrophages, Pneumocystis carinii pneumonitis (PCP) could be detected in 100% of cases. Transbronchial biopsy permitted the correct diagnosis in only 65.2% of cases. Retrospective analysis of slides is possible after a long period as no significant loss of fluorescence occurs after 4 years. Thus fluorescence microscopy permits the diagnosis of Pneumocystis carinii without any additional staining or loss of time.     

Morphological study of bacteria of the respiratory system using fluorescence microscopy of Papanicolaou-stained smears with special regard to the identification of Mycobacteria sp.

In Papanicolaou-stained smears certain structures such as nucleoli, Pneumocystis carinii , Charcot-Leyden crystals, bacteria and fungi show a brilliant fluorescence. the morphological characteristics of microorganisms which can be detected by this system, especially mycobacteria, are described. This screening method offers the possibility of providing the clinician with a provisional diagnosis within hours. Proof of the nature of the organisms should be obtained by culture.     

Identification of Phagocytosed Pneumocystis Carinii In Human Pulmonary Alveolar Macrophages

Examination of Papanicolaou-stained bronchoalveolar lavage samples from cases with Pneumocystis carinii pneumonitis under ultra-violet light reveals alveolar macrophages packed with fluorescent inclusions. Immunoenzymatic staining of the alveolar macrophages with a monoclonal antibody specific for P. carinii (3F6) showed that these inclusions contain intact pneumocysts or their degradation products. Fluorescence microscopy of Papanicolaou-stained smears is advocated as a sensitive and specific method of diagnosing P. carinii infection.     

Fine needle aspiration (FNA)-induced necrosis of tumours of the thyroid

Two cases of necrosis of thyroid oxyphilic tumours following FNA are reported. the first patient received surgery 4 weeks after FNA and histological examination revealed an encapsulated and totally necrotic tumour 2 cm in diameter. In the second patient surgery was performed after 25 days. Histological examination showed a 0.7 cm diameter tumour consisting mainly of fibrous tissue with residual oxyphilic tumour cells only in a small peripheral rim. In both patients no capsular or vascular invasion and no blood vessel thrombosis were present. A review of the literature revealed that oxyphilic tumours are susceptible to post-FNA necrosis, which might be due to the compromised vascular supply after FNA in conjunction with the intrinsic energy deficiency of oncocytic tumour cells.