TABLE 1.

Comparison of different in situ hybridization protocols for miRNA detection.

Reference	Protease/prehybrization	Probe label	Probe concentration	Hybridization time/temperature	Wash	TSA	Tissue	Colabel/RT in situ PCR
13	Yes/Yes	FITC	1.2 pmol per 20 μl	2 h/53 °C	53 °C, 0.1× SCC	Yes	FFPE	No/No
15	No/Yes	FITC Digoxigenin	1.0 pmol per 20 μl	1 h/53 °C	55 °C, 0.1× SCC	Yes	Frozen	No/No
16	No/Yes	Digoxigenin	3.0 pmol per test	1 h/55 °C	55 °C, 0.1× SCC	Yes	Frozen	No/No
12	Yes/Yes	Digoxigenin	0.1 pmol per 20 μl	15 h/50 °C	60 °C, 0.1× SCC	No	Frozen	Yes/No
11	No/Yes	FITC	Not listed	3 h/37 °C	37 °C, not listed	No	FFPE	No/No
This protocol	Yes/No	Digoxigenin	2−40 pmol per 20 μl	15 h/37 °C	4 °C, 0.1× SCC	No	FFPE	Yes/Yes

FITC, fluorescein isothiocyanate; FFPE, formalin-fixed, paraffin-embedded tissue; TSA, tyramide signal amplification.