Table 1.

Enzymatic dissociation strategies and applications of nuclei from FFPE tissues.

Tissues	Thickness	Protease	Time	Application	References
tumor	30 μm	0.5% pepsin	30 min	DNA content	[17]
brain, lung, breast, testis, kidney, and colon tumor	50 μm	0.5% pepsin	30 min	ISH	[18]
——	50 μm	0.5% pepsin	30 min	FISH	[19]
brain	——	0.5% pepsin	30 min	FISH	[21]
breast, ovary, tumor	40 μm	0.005% pepsin	2 h	FISH	[22]
tumor	——	0.25% trypsin	Overnight	DNA ploidy	[23]
——	20–30 μm	0.005% PK	30 min	FISH	[20]
HNSCC	5 μm	PK	1 h	FISH	[24]
lymph nodes, tonsils	——	0.01% PK	2 h	ISH, FISH	[27]
liver cancer	10 μm	0.005% PK	2 h	FISH	[28]
breast, brain, bladder, ovarian, and pancreas	40–60 μm	collagenase, hyaluronidase	Overnight	aCGH, WES	[25]
breast cancer	100 μm		16 h	sn-WGSA	[26]

PK: proteinase K; ISH: in situ hybridization; FISH: fluorescence in situ hybridization; aCGH: array comparative genomic hybridization; WES: whole-exome sequencing; sn-WGSA: single-nucleus whole-genome DNA copy number; HNSCC: head and neck squamous cell carcinoma.