Zeiss LSM 510 META
The LSM 510 META is located in a DCML satellite facility in Biomed 247-2 in the laboratory of Brian Storrie. The DCML's LSM 510 META is an inverted microscope (Axiovert 200), and is equipped with heated stage chamber and objective warmer from PECON for live cell experiments. It has two PMTs, a detector for bright field image acquisition and a polychromatic detector named the META detector (see below). It has a 100 watt mercury lamp and filter sets to view via oculars the fluorescence of DAPI, green, or red fluorophores in conventional widefield microscopy mode. Movement of the stage is not motorized in the XY direction; however, the DCML's Zeiss LSM 410 does have a motorized XY stage. Technical data about the LSM 510 META can be found at the Zeiss website by clicking this link.
Reserving the LSM 510 META
Priority for use of the instrument is given to several "primary" user laboratories who submitted a shared instrument grant (SIG) to obtain the microscope; these users are sometimes referred to as "SIG" users of the instrument. They are Giulia Baldini, Richard Kurten, Vladimir Lupashin, Khaled Machaca, Robert Safirstein, Brian Storrie, Jerry Ware, and BRIN/INBRE students (Lawrence Cornett). Someone other than a SIG user, an outside user, can use the LSM 510 META whenever it is available if they have been trained to use the instrument, or with assistance by DCML staff. Please contact DCML staff for more details about SIG users and scheduling the LSM 510 META. DCML staff would prefer that the LSM 510 be used by individuals needing capabilities not provided by the LSM 410 (see below).
Basic Capabilities of LSM 510 META Not Available on LSM 410
	Precise Laser Placement: Laser illumination can be accurately controlled permitting localized photo-activation/uncaging, irregular object photo-bleaching/uncaging, rapid line scan, and even irregular region of interest (ROI) studies.
	Multitracking: Multiple scanning set-ups or tracks are executed automatically in rapid sequence, and the acquired digital images are automatically overlaid; consequently, simultaneous excitation of fluorophores with considerable spectral overlap or bleed through can be avoided. While acquiring z-stack or time lapse image sequences, all tracks can be sequentially scanned at each z-position or time point. Although the LSM 410 is not capable of automatic multitracking while acquring time lapse or z-stack images, macros or "plug-ins" can be written and installed to allow this in the future.
	META Detector: Light emitted from a fluorescent sample is separated into various wavelengths with a grating (prism), and an array of 32 detector elements, collectively referred to as the META detector, samples the light in 10.7nm increments. Each detector can be electronically switched on or off, allowing the microscopist to choose wavelengths of light detected and displayed in digital images. Analogous to creating custom band pass filters, any number of consecutive detectors can be activated for each track of a multitracking scenario. The META detector can also be used to acquire Lambda Stacks. The LSM software displays the images acquired from the META detector channels as an image series from shortest to longest wavelength; this series of images can be overlaid to create a "stack" of images called a Lambda Stack.
	Emission Fingerprinting and Linear Unmixing: By a process called emission fingerprinting, ROIs are selected from the Lambda Stacks acquired from samples with a single fluorophore, or a region of an image stack representing only one fluorophore, and the LSM software creates and stores spectral curves from these ROIs. These spectral data are then utilized by the LSM software to distinguish two or more fluorophores within the same sample (ie. within the same pixel of the image) even if they have considerable spectral overlap (eg. EGFP and EYFP or even EGFP and FITC). This spectral separation technique is called linear unmixing. To see descriptions of emission fingerprinting and linear unmixing that include figures and fluorescence images, click the Zeiss Emission Fingerprinting web link or the web link for the Biotechniques article about linear unmixing.
Zeiss LSM Image Browser
A free LSM Image Browser program for viewing images captured on the LSM 510 META and managing image databases in MDB format can be downloaded from the Carl Zeiss website. Zeiss Axiovision LE software, free to download from Zeiss, is also capable of reading images captured by the LSM 510, including z-stacks and time series, but it does not support Lambda stacks.
Objective Lenses
	10x/0.45 Plan Apochromat
	20x/0.75 Plan Apochromat
	40x/1.3 Plan Neofluar oil DIC
	63x/1.4 Plan Apochromat oil DIC
	40x/1.2 C-Apochromat water-immersion DIC, cover glass thickness correction collar
Lasers
	Diode 405nm, 30mW
	Ar/ML 458/477/488/514nm, 30mW
	DPSS 561nm, 10mW
	HeNe 633nm, 5mW