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<p><strong>Objective</strong> To explore the value of detecting early secretory antigenic target-6 (ESAT-6) in cerebrospinal fluid (CSF) monocytes by laser scanning confocal microscopy (LSCM) techniques in early diagnosis of tuberculous meningitis (TBM), and to find a more specific method for early diagnosis.  <strong>Methods</strong> Double immunofluorescence staining was applied to detect ESAT-6, and LSCM was used to observe the dyeing result of CSF monocytes and to analyze three-dimensional images. ESAT-6 positive cells will present red fluorescence cytoplasm and blue fluorescence nuclei, and negative cells present blue fluorescence nuclei while the cytoplasm is not stained.  <strong>Results</strong> ESAT-6 was mainly expressed with red fluorescence in cytoplasm of CSF monocytes in patients with TBM. Among 35 cases of TBM patients, there were 28 patients (80%) ESAT-6 positive, one case (2.86%) ESAT-6 positive in the control group, and the difference was statistically significant (<em>χ²</em> = 42.918, <em>P</em> = 0.000). ESAT-6 sensitivity was 80% and specificity was 97.14%. <strong> Conclusions</strong> The detection of ESAT-6 can provide a certain basis for the early diagnosis of TBM and has high specificity. LSCM techniques can realize simultaneously the observation of multiple fluorescence and provide clear tomography and three-dimensional images. The cytological research has been improved to a new level.</p><p> </p><p><strong>DOI: </strong>10.3969/j.issn.1672-6731.2016.08.010</p>