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PACMan: A software package for automated single-cell chlorophyll fluorometry.
Pontén, Olle; Xiao, Linhong; Kutter, Jeanne; Cui, Yuan; Wählby, Carolina; Behrendt, Lars.
Affiliation
  • Pontén O; Department of Organismal Biology, Science for Life Laboratory, Uppsala University, Uppsala, Sweden.
  • Xiao L; School of Mathematics & Statistics, The University of Melbourne, Parkville, VIC, Australia.
  • Kutter J; Department of Organismal Biology, Science for Life Laboratory, Uppsala University, Uppsala, Sweden.
  • Cui Y; Department of Organismal Biology, Science for Life Laboratory, Uppsala University, Uppsala, Sweden.
  • Wählby C; Department of Organismal Biology, Science for Life Laboratory, Uppsala University, Uppsala, Sweden.
  • Behrendt L; Department of Information Technology and Science for Life Laboratory, Uppsala University, Uppsala, Sweden.
Cytometry A ; 105(3): 203-213, 2024 03.
Article in En | MEDLINE | ID: mdl-37864330
ABSTRACT
Microalgae, small photosynthetic unicells, are of great interest to ecology, ecotoxicology and biotechnology and there is a growing need to investigate the ability of cells to photosynthesize under variable conditions. Current strategies involve hand-operated pulse-amplitude-modulated (PAM) chlorophyll fluorimeters, which can provide detailed insights into the photophysiology of entire populations- or individual cells of microalgae but are typically limited in their throughput. To increase the throughput of a commercially available MICROSCOPY-PAM system, we present the PAM Automation Control Manager ('PACMan'), an open-source Python software package that automates image acquisition, microscopy stage control and the triggering of external hardware components. PACMan comes with a user-friendly graphical user interface and is released together with a stand-alone tool (PAMalysis) for the automated calculation of per-cell maximum quantum efficiencies (= Fv /Fm ). Using these two software packages, we successfully tracked the photophysiology of >1000 individual cells of green algae (Chlamydomonas reinhardtii) and dinoflagellates (genus Symbiodiniaceae) within custom-made microfluidic devices. Compared to the manual operation of MICROSCOPY-PAM systems, this represents a 10-fold increase in throughput. During experiments, PACMan coordinated the movement of the microscope stage and triggered the MICROSCOPY-PAM system to repeatedly capture high-quality image data across multiple positions. Finally, we analyzed single-cell Fv /Fm with the manufacturer-supplied software and PAMalysis, demonstrating a median difference <0.5% between both methods. We foresee that PACMan, and its auxiliary software package will help increase the experimental throughput in a range of microalgae studies currently relying on hand-operated MICROSCOPY-PAM technologies.
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Full text: 1 Database: MEDLINE Main subject: Dinoflagellida / Microalgae Language: En Journal: Cytometry A Year: 2024 Type: Article Affiliation country: Sweden

Full text: 1 Database: MEDLINE Main subject: Dinoflagellida / Microalgae Language: En Journal: Cytometry A Year: 2024 Type: Article Affiliation country: Sweden