Fluorescence microscopy of mycobacteria in pleural effusions.

The diagnostic advantage of fluorescence microscopy (FM) of Papanicolaou-stained cytological specimens obtained by bronchoscopy has been described previously. This study was designed to evaluate the method's diagnostic benefit in cytological preparations of pleural effusions in cases of active pulmonary tuberculosis. In contrast to bronchial material there is no advantage in cytological evaluation of pleural effusions by FM.

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.

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.

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.

The cytodiagnosis of meningeal involvement in familial haemophagocytic lymphohistiocytosis.

The cytological appearances of the cerebrospinal fluid (CSF) in two cases of familial haemophagocytic lymphohistiocytosis (FHL) are described. The presence of numerous lymphocytes and immature macrophages in the CSF, some of which showed lymphohistiocytosis, was indicative of meningeal involvement. The appearance of large numbers of immature macrophages indicated rapid deterioration and death within a few weeks. Furthermore, some CSF samples taken at times when the patients were asymptomatic contained suspicious cells, indicating asymptomatic persistence of the meningeal lesions. Thus, cytological examination of CSF can assist with the management of patients with this rare disease.

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.

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.

Quantitative differences in phagocytosis and degradation of Pneumocystis carinii by alveolar macrophages in AIDS and non-HIV patients in vivo.

Bronchoalveolar lavage (BAL) specimens (n = 213) from AIDS and non-HIV immunosuppressed patients were investigated for the presence of Pneumocystis carinii infection by fluorescence microscopy of Papanicolaou-stained slides. Alveolar casts, extracellular pneumocysts and phagocytosed cysts and their degradation products in pulmonary alveolar macrophages were identified. The number of phagocytosed pneumocysts within human pulmonary alveolar macrophages was recorded and correlated with the number of extracellular cysts and alveolar casts, in both groups of patients. Both phagocytic and degradation capacity were depressed in AIDS patients. This observation may explain the large number of extracellular organisms found in BAL specimens of AIDS patients compared with non-HIV-positive immunocompromised individuals.

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.

[Fine needle puncture cytology in metastatic hepatocellular carcinoma]. / Feinnadelpunktionszytologie bei metastasierendem hepatozellulärem Karzinom.

Distant metastases occur in 50% of cases of HCC. Bones are involved in 11% with the spine as the most important localization. In the reported case a tumorous mass in the liver, suspicious for HCC according to clinical and cytological criteria, was proved to be malignant by the cytologic examination of a metastasis to the sacrum.

Nonspecific esterase activity of human alveolar macrophages in routine cytology.

Alpha-naphthyl acetate esterase (ANAE) activity in alveolar macrophages was demonstrated in air-dried smears from samples of 82 sputa, 47 bronchial secretions and 14 bronchial lavages from 113 patients. Enzyme activity was estimated by a semiquantitative scoring method. There was a 7.4% loss of activity after 24 hours of storing the unfixed material, increasing to 14.4% after three days, while storing the air-dried smears for up to four weeks did not change the ANAE activity. The mean cellular esterase activity was correlated to the clinical and cytologic findings. A stringent correlation could be found for patients smoking more than 30 cigarettes a day; they had a 17% increase in activity as compared to nonsmokers. In patients with bronchial asthma, the activity was 18% higher than the total mean. In three patients with pulmonary embolisms, the ANAE activity was also increased. Treatment with a combination of cytostatics and a corticoid caused a severe decrease. No correlation could be found to age, sex, inflammation or malignant or cardiac diseases. These findings indicate that the application of the ANAE reaction to routine cytologic specimens can contribute to the functional characterization of human alveolar macrophages.

Peroxidase activity in human alveolar macrophages demonstrated by 3-amino-9-ethyl-carbazol as substrate.

Airdried smears of routine sputa, bronchial secretions, and bronchus lavages were incubated to demonstrate peroxidase activity in the alveolar macrophages, with 3-amino-9-ethyl-carbazol as substrate. In light microscopic examination two deposition patterns of reaction product could be found: Reaction product only to be found on, resp. in the nuclear membrane; Reaction product deposited in granules throughout the cytoplasm, some cells being totally loaded. However, most of the alveolar macrophages did not show any peroxidase activity. Thus the population of alveolar macrophages comprises resident and exudate forms.