Keck Jonathon, Chambers James P, Forsthuber Thomas, Gupta Rishein, Arulanandam Bernard P
South Texas Center for Emerging Infectious Diseases, University of Texas at San Antonio, One UTSA Circle, San Antonio, TX, 78249, United States of America.
MethodsX. 2019 Aug 30;6:1925-1932. doi: 10.1016/j.mex.2019.08.005. eCollection 2019.
Although manual enumeration of inclusion forming units is the most widely accepted means of quantification in the field, it is both time consuming and subject to inherent investigator bias. We report here a rapid, ., minutes . hours, modified automated Fluorospot means of assessment that is linear (<1200 dots per well). Because the Fluorospot enumerated tissue culture plate/well can also be quantified using traditional manual counting, newly derived Fluorospot data can easily be compared to previously established manual enumeration data requiring no new reference norms. •Concurrent enumeration of chlamydial IFU using automated and manual methods of counting on same tissue culture plate.•Rapid method of counting chlamydial IFU reducing time from hours to minutes.
尽管手动计数包涵体形成单位是该领域最广泛接受的定量方法,但它既耗时又存在调查人员固有的偏差。我们在此报告一种快速(数分钟而非数小时)、经过改进的自动化荧光斑点评估方法,该方法具有线性关系(每孔<1200个点)。由于使用传统手动计数也可以对荧光斑点计数的组织培养板/孔进行定量,因此新获得的荧光斑点数据可以轻松地与先前建立的手动计数数据进行比较,无需新的参考标准。•在同一组织培养板上使用自动和手动计数方法同时计数衣原体包涵体形成单位。•快速计数衣原体包涵体形成单位的方法,将时间从数小时缩短至数分钟。