Table 1 Methods for identifying... | American Society of Hematology

Table 1

Methods for identifying hematopoietic cells in zebrafish

Cell type	Histochemical stain(s)	Flow cytometry⁵⁹	Selection of riboprobe markers for whole mount in situ hybridization*†		Antibody	Transgenic fluorescent strain
Cell type	Histochemical stain(s)	Flow cytometry⁵⁹	Transcriptional marker(s)	Lineage marker(s)	Antibody	Transgenic fluorescent strain
Hemangioblast/stem cell	—	FSC^intermediate SSC^low	*tal1(scl)^88,89*	—	—	tal1:EGFP^49,156
		Hoechst dye efflux¹⁵⁴	hhex¹⁵⁵
Definitive hematopoiesis onset	—	—	*runx1⁴⁸*	—	—	cd41:EGFP^low⁶²
			cmyb⁴⁴			lmo2:EGFP⁹⁷
						lmo2:dsRED⁹⁷
			lmo2⁴⁴
Erythrocyte	o-Dianisidine⁴¹	FSC^low SSC^high	gata1⁴¹	Globins⁶⁸	β-spectrin¹²²	gata1:EGFP¹⁵⁹
	Diaminofluorene⁸		klf4^85,157	alas2¹²⁵
			znfl2⁹³	jak2a¹¹²		gata1:dsRED¹⁶⁰
				fech¹⁵⁸
Early myeloid cell	—	—	*spi1(pu.1)³⁸*	—	spi1⁴³	spi1:EGFP^58,161
						fli1a:GFP¹⁶²
Neutrophil	Myeloperoxidase^51,52	FSC^high SSC^high	cebpa¹⁶³	*mpx^51,52*	mpx¹⁶⁴	mpx:EGFP^61,136
	Sudan black⁴³		mych²⁸			CLGY463:YFP²⁸
						CLGY383:YFP²⁸
Monocyte/macrophage	Neutral red⁴²	FSC^high SSC^high	drl⁴⁰	*csf1r¹⁰¹*	WCL15¹⁴⁵⁹
Myelomonocytic‡	—	FSC^high SSC^high	—	lcp1(l-plastin)⁴⁰	l-plastin^42,72	lyz:EGFP¹⁴⁴
				lyz¹⁶⁵
				mmp9¹⁶⁶		lyz:dsRED¹⁴⁴
				ncf1¹⁶⁷
Eosinophil	Periodic acid-Schiff⁵¹	FSC^high SSC^high	—		—	gata2:EGFP^59,168
T lymphocyte	—	FSC^low SSC^low	ikzf1(ikaros)⁷⁵	*rag1⁷⁴*	—	lck:EGFP⁶⁰
				rag2⁷⁴		rag2:EGFP⁶⁰
						rag2:dsRED¹³⁰
B lymphocyte	—	FSC^low SSC^low		*ighm⁷⁶*	—	—
Thrombocyte	—	FSC^low SSC^low	zfpm1(fog1)⁹⁸	*itga2b(CD41)⁶²*	GPIIb/IIIa⁸⁰	CD41:EGFP⁶²
				mpll⁶²

Cell type	Histochemical stain(s)	Flow cytometry⁵⁹	Selection of riboprobe markers for whole mount in situ hybridization*†		Antibody	Transgenic fluorescent strain
Cell type	Histochemical stain(s)	Flow cytometry⁵⁹	Transcriptional marker(s)	Lineage marker(s)	Antibody	Transgenic fluorescent strain
Hemangioblast/stem cell	—	FSC^intermediate SSC^low	*tal1(scl)^88,89*	—	—	tal1:EGFP^49,156
		Hoechst dye efflux¹⁵⁴	hhex¹⁵⁵
Definitive hematopoiesis onset	—	—	*runx1⁴⁸*	—	—	cd41:EGFP^low⁶²
			cmyb⁴⁴			lmo2:EGFP⁹⁷
						lmo2:dsRED⁹⁷
			lmo2⁴⁴
Erythrocyte	o-Dianisidine⁴¹	FSC^low SSC^high	gata1⁴¹	Globins⁶⁸	β-spectrin¹²²	gata1:EGFP¹⁵⁹
	Diaminofluorene⁸		klf4^85,157	alas2¹²⁵
			znfl2⁹³	jak2a¹¹²		gata1:dsRED¹⁶⁰
				fech¹⁵⁸
Early myeloid cell	—	—	*spi1(pu.1)³⁸*	—	spi1⁴³	spi1:EGFP^58,161
						fli1a:GFP¹⁶²
Neutrophil	Myeloperoxidase^51,52	FSC^high SSC^high	cebpa¹⁶³	*mpx^51,52*	mpx¹⁶⁴	mpx:EGFP^61,136
	Sudan black⁴³		mych²⁸			CLGY463:YFP²⁸
						CLGY383:YFP²⁸
Monocyte/macrophage	Neutral red⁴²	FSC^high SSC^high	drl⁴⁰	*csf1r¹⁰¹*	WCL15¹⁴⁵⁹
Myelomonocytic‡	—	FSC^high SSC^high	—	lcp1(l-plastin)⁴⁰	l-plastin^42,72	lyz:EGFP¹⁴⁴
				lyz¹⁶⁵
				mmp9¹⁶⁶		lyz:dsRED¹⁴⁴
				ncf1¹⁶⁷
Eosinophil	Periodic acid-Schiff⁵¹	FSC^high SSC^high	—		—	gata2:EGFP^59,168
T lymphocyte	—	FSC^low SSC^low	ikzf1(ikaros)⁷⁵	*rag1⁷⁴*	—	lck:EGFP⁶⁰
				rag2⁷⁴		rag2:EGFP⁶⁰
						rag2:dsRED¹³⁰
B lymphocyte	—	FSC^low SSC^low		*ighm⁷⁶*	—	—
Thrombocyte	—	FSC^low SSC^low	zfpm1(fog1)⁹⁸	*itga2b(CD41)⁶²*	GPIIb/IIIa⁸⁰	CD41:EGFP⁶²
				mpll⁶²

FSC indicates forward scatter; SSC, side scatter; —, not applicable.

More details and a comprehensive literature listing about individual genes and strains can be found on the Zebrafish Information Network (ZFIN).¹⁶⁹

†

Most specific marker in bold.

‡

Precise leukocyte lineage specificity uncertain.

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