QClus: a droplet filtering algorithm for enhanced snRNA-seq data quality in challenging samples
: Schmauch, Eloi; Ojanen, Johannes; Galani, Kyriakitsa; Jalkanen, Juho; Harju, Kristiina; Hollmén, Maija; Kokki, Hannu; Gunn, Jarmo; Halonen, Jari; Hartikainen, Juha; Kiviniemi, Tuomas; Tavi, Pasi; Kaikkonen, Minna U; Kellis, Manolis; Linna-Kuosmanen, Suvi
Publisher: OXFORD UNIV PRESS
: OXFORD
: 2025
: Nucleic Acids Research
: NUCLEIC ACIDS RESEARCH
: NUCLEIC ACIDS RES
: 53
: 1
: 13
: 0305-1048
: 1362-4962
DOI: https://doi.org/10.1093/nar/gkae1145
: https://doi.org/10.1093%2Fnar%2Fgkae1145
: https://research.utu.fi/converis/portal/detail/Publication/477906384
Single-nuclei RNA sequencing remains a challenge for many human tissues, as incomplete removal of background signal masks cell-type-specific signals and interferes with downstream analyses. Here, we present Quality Clustering (QClus), a droplet filtering algorithm targeted toward challenging samples. QClus uses additional metrics, such as cell-type-specific marker gene expression, to cluster nuclei and filter empty and highly contaminated droplets, providing reliable filtering of samples with varying number of nuclei and contamination levels. In a benchmarking analysis against seven alternative methods across six datasets, consisting of 252 samples and over 1.9 million nuclei, QClus achieved the highest quality in the greatest number of samples over all evaluated quality metrics and recorded no processing failures, while robustly retaining numbers of nuclei within the expected range. QClus combines high quality, automation and robustness with flexibility and user-adjustability, catering to diverse experimental needs and datasets.
:
Aarne Koskelo Foundation [to E.S. and S.L.K.]; Academy of Finland [333021 to M.U.K and 342074 to S.L.K.]; Antti and Tyyne Soininen Foundation [to E.S.]; University of Eastern Finland [to E.S.]; Finnish Cultural Foundation [to E.S.] Finnish Foundation for Cardiovascular Research [to M.U.K. and S.L.K.]; Maud Kuistila Memorial Foundation [to S.L.K.]; Orion Research Foundation [to E.S. and S.L.K.]; Saastamoinen Foundation [to E.S.]; Sigrid Juselius Foundation [to M.U.K. and S.L.K.]; Väisälä Fund [to E.S.]; Yrjö Jahnsson Foundation [to E.S. and S.L.K.]. Funding for open access charge: The Academy of Finland.
