FujiFilm LAS-3OOO More sensitivity for chemiluminescent detection, Digitizing functions

Luminescent image analyzer (LAS) for applications requiring high

sensitivity and a wide range of fluorescent capabilities

GFP expression in tobacco leaf

Detection of EPO as doping substance in sports Isoelectric profiles of natural EPO found in normal human urine (B) and the bands found in urine after injections of epoetin or darbepoetin.

Data courtesy of Dr. Françoise Lasne.

Laboratoire National de Dépistage du Dopage,

Châtenay-Malabry, France

The LAS-3000 imaging system combines Fujifilm’s high-sensitivity Super CCD camera technology with the added ver- satility of white, blue, green and red EPI illuminators. The Super CCD imaging chip, binning mode and specially designed camera lens allow researchers to capture faint-light luminescent images with unprecedented sensitivity and resolution. Multicolor illuminator options enlarge application area in fluorescent imaging.

More sensitivity for chemiluminescent detection

Western blotting, Southern blotting and Northern blotting detection by chemilumi- nescence is a widely accepted method. The use of a cooled CCD camera system enables the generation of a digital image and quanti- tative analysis of the image’s signal strength.

Several state-of-the-art technologies were incorporated to make the LAS-3000 system as sensitive as the conventional film method:

•The newly designed F0.85 high-sensitiv- ity Fujinon lens gathers as much light as possible onto the CCD.

•The CCD is a 3.2M-pixel Super CCD having large octagonal-shaped CCD pixels 10.75 x 10.75µm in size.

•The CCD is cooled to -30°C by peltier when the environment is below 28°C.

•Pixel binning in the LAS-3000 electroni- cally increases the pixel area to increase sensitivity. The binning mode includes four binning levels: 1 x 2 (Standard),

2 x 4 (High), 4 x 8 (Super) and 8 x 16 (Ultra). In the Ultra mode, the exposure time is nearly 60 times faster than Standard. An image smoothing function increases the pixels to make the image size the same as the Standard.

•Long exposure times are required to capture images of extremely low-light samples. The low-noise design of the LAS-3000 system includes default settings for a two-hour exposure and a 15-hour exposure in addition to manual settings for exposures as long as 30 hours.

•A non-parallax (NP) tray is used to elimi- nate the parallax effect when imaging chemiluminescence in the 96-well plate. Up to two plates can be placed.

More versatility for fluorescent detection

More and more fluorescent dyes are intro- duced into the biochemistry and molecular biology fields for staining proteins and nucleic acids in gels, membranes and well plates. Expression analysis using green fluo- rescent protein (GFP) and red fluorescent protein (RFP) can be detected by blue exci- tation or green excitation respectively.

The LAS-3000 includes multiple LED options for various fluorescence applications, including blue LED (470nm), green LED (520nm) and red LED (630nm) for epi-illu- mination and a UV (312nm) light source for DIA-illumination.

The five-position filter turret holds 77mm filters, such as Y515 (general purpose for blue LED excitation), 510DF10 (GFP, etc.), 575DF20 (RFP, Cy™3, Pro-Q™Diamond, etc.), R670 (Cy™5, ALEXA Fluor® 633, etc.) and 605DF40 (EtBr, etc.).

Digitizing functions

Capturing images as we see them is called digitizing. For CBB-stained gel or silver- stained gel, use of the white LED DIA- illuminator at iris f2.8 without any filter is recommended. For digitizing the molecular weight standard on blotted membrane, white light epi-illuminator, which is located in the center part of the blue, green and red LED epi-illuminator unit, is used.

Easy-to-use operation

Samples are placed on the appropriate tray and placed in the intelligent dark box. After closing the door, all the functions are controlled remotely from the Image Reader software. The software automatically rec- ognizes the type of illuminator installed or changed during operation.

Users are able to control the LAS-3000 with the Image Reader software in either the user-friendly Lite mode or the more advanced Pro mode. The modes are selected with the click of a mouse.

Super CCD - By rotating pixels 45 degrees to form an interwoven layout, the Super CCD’s pixel pitch in the horizontal and vertical directions is narrower than in the diagonal direction, achieving higher hori- zontal and vertical resolution.

Quantitative image analysis function

The 16-bit signal accumulated on each CCD pixel is processed through an A/D converter to form a digital image. The image data obtained at the closed shutter condition is the dark frame. The image data of an even, flat sample is the flat frame. To analyze the image data, the dark frame and flat frame data are subtracted from the original image. This correc- tion function is included in the Image Reader software. MultiGauge software for Windows® and ImageGauge software for Mac™ are the standard image analysis software included in the ScienceLab software provided by Fujifilm.

Fujifilm Super CCD Area

Type imaging chip

FUJINON lens VRF43LMD and

five-position filter turret

Applications

Fluorescence/Chemifluorescence

Blue:	SYBR® Green I & II	SYBR® Gold
	SYPRO® Ruby	SYPRO® Orange
	SYPRO® Tangerine	FITC
	FAM™	EGFP
	ECFP	AttoPhos™
	DY-458XL
Green:	SYPRO® Red	Cy™3
	TAMRA™	ROX™
	HEX™	Alexa Fluor® 532
	Alexa Fluor® 546	Deep Purple
	Pro-Q™ Diamond	Rhodamine Red™
	BODIPY®576/589	NED™
	R-phycoerythrin	RFP
	HNPP	DY-520XL
	DY-547
Red:	Alexa Fluor® 633	Alexa Fluor® 635
	Alexa Fluor® 647	Cy™5
	BODIPY®650/665	DiD
	TOTO®-3	DDAO phosphate
	DY-647
UV:	Ethidium Bromide	SYPRO® Rose
White:	Silver stained	CBB
	NBT / BCIP	X-ray film

Chemiluminecence

ECL™	ECL Plus™
ECL Advance™	Lumi-Light Plus
®

NP Tray	The five filter options available	LED units
	for the LAS-3000.

Super Signal	CDP-Star™
CSPD®	Renaissance™
Bright-Star™

C H E M I L U M I N E S C E N C E • F L U O R E S C E N C E / C H E M I F L U O R E S C E N C E • D I G I T I Z A T I O N