Table 3.

Features of several fluorescence microscopy techniques in the light of fast imaging. The normalized dwell time is the ratio of the time a pixel is illuminated divided by the total time it takes to acquire one frame. The ratio of the collection volume to the illumination volume gives a good feel of how the system will behave in terms of bleaching. Struct. Illum.: Structured Illumination, Pt: single beam point scanner, Multipt: multi-beam, 1D: line array detector, 2D: two-dimensional array detector (CCD,…), P: point detector (PMT,…), y: yes, n: no, Pin: adjustable slit, Sli: adjustable width slit. The more  ^* the better. †: requires additional setup.  ^‡: in one direction only.

Microscope type	Illumination type	Normalized dwell time	Detector type	$\frac{\text{Collection}\text{volume}}{\text{Illumination}\text{volume}}$	Sectioning capability	Penetration depth	Region of interest	Adjustable collection volume	Conventional mounting	Lateral resolution	Temporal resolution	Bleaching	Reference
Widefield
Conventional	Uniform	1	2D	*	*	*	n	∅	y	**	***	*	(Murray et al., 2007)
Struct. illum. sectioning	Patterned	1	2D	*	**	*	n	∅	y	***	*	*	(Neil et al., 1997)
Confocal
Single beam	Pt	10−6	P	**	***	**	y	Pin	y	****	**	**	(Wilson, 1990)
Spinning disk	Multipt	10−3	2D	**	***	**	y†	∅	y	***	***	**	(Graf et al., 2005)
Line scanning	Line	10−3	1D	**	***	**	y‡	Sli	y	***	****	**	(Wolleschensky et al., 2006)
Multiphoton
Single beam	Pt	10−6	P	***	***	***	y	∅	y	***	***	***	(Helmchen and Denk, 2005)
Multi-beam	Multipt	10−5	2D	***	***	***	y	∅	y	***	***	***	(Niesner et al., 2007)
SPIM	Sheet	1	2D	***	****	***	n	∅	n	***	***	***	(Huisken et al., 2004)