Statement of Work Fluorescent Microscope RFQ.pdf

Project Acquisition of the fluorescent microscope for the precision-guided Sterile Insect Technique (pgSIT) project at IMMDL

1.0 Scope of Work

For the generation of transgenic strains, one or multiple of these photoproteins are usually engineered into the genome as the transgenic markers. The fluorescent microscope provides multiple sources of illumination including LED and UV.

Various filters can modify the UV spectra to emit optimal wavelengths to visualize photoproteins, such as GFP, BFP, DsRED and mCherry. Therefore, this unit critically enables us to screen the transgene-containing fly individuals and evaluate levels of their activity or effect after the microinjection procedure is successfully finished.

2.0 Background

The Insect Management and Molecular Diagnostics Laboratory (IMMDL) is responsible for developing and validating technologies used to detect and diagnose plant pests. This laboratory also identifies, develops, and provides technology transfer of a wide range of scientific methods to PPQ and State Departments of Agriculture – geographic information systems, biological control, area-wide pest management, and sterile insect technology (SIT) support for the Mexican fruit fly (Mexfly) eradication program. For enhancing the Mexfly control, the frontier genetic approaches using CRISPR technology are highly demanded to be implemented. And it requires the generation of genetically modified (GE) or transgenic strains using the recombinant DNA cassettes that contain fluorescent markers. Thus, the fluorescent microscope is prerequisite for verifying and maintaining the individual GE lines in the laboratory.

3.0 Technical Requirements

The acquisition of the fluorescent microscope is necessary for the generation of transgenic mosquito strains. The performance must be high enough to succeed many critical microinjection processes with full satisfaction in insect transgenesis.

Specifications of this requirement include:

Fluorescent microscope:

• Includes multiple sources of illumination (LED and UV)

• Interpupillary distance range of 57-63mm

• Observation angle = 30°

• Compatible with multiple filters that modify the UV spectra to emit optimal wavelengths to visualize photoproteins, such as GFP, BFP, DsRED and mCherry

• Supports live specimen viewing

• Required accessories:

Optics carrier

• Minimum magnification range: 7.3x-120x (1.0x objective, 10x eyepieces)

• Click stop zoom settings

• Separate illumination beam path for fluorescence

• Minimum of four position fluorescence filter turrets

• Includes magnification, fluorescence filter, and iris diaphragm functions

• Apochromatic corrected optics

• Parfocal plan-apochromat objectives in at least four magnifications between 0.6x-3x

• Compatible objective threading

LED light source

• Generates white light output (390-680 nm) through the combination of multiple solid state light outputs

• Mercury-free

• Supports fluorescence and stereo microscopy applications

• As maintenance-free as possible

• Extended lifetime while maintaining lamp quality

• Can be triggered to synchronize illumination with camera exposure and minimize photobleaching

Filter sets

• Green Fluorescent Protein (GFP) minimums: excitation 470nm at 40x; emission 525nm at 50x

• mCherry Fluorescent Protein minimums: excitation 560nm at 40x; emission 630 at 75x

• TagBFP minimums: excitation 405nm at 20x; emission 460nm at 50x

LED Spotlight Illuminator

• LEDs at the end of 400+ mm flexible arms (minimum of 2 arms)

• Microscope provides power through illuminator’s integrated cable(s)

• Control via microscope software and physical control keypad

Trinocular tube

• Allows 100% switch between camera and eyepieces

Microscope Camera

• Complementary metal oxide semiconductor (CMOS) image sensors

• Maximum pixel size 2.4 µm x 2.4 µm

• Minimum imaging size: 3072 x 2048 pixels

• RGB color filter

• Color depth: 3x12 bits

• Maximum exposure time: 1 second

• Minimum exposure time: 1 millisecond or less

• Minimum frames per second video: 15

• Minimum dynamic range: 72dB

• Gain 1x-251x

• Compatible with Windows operating system

• Includes appropriate mounting hardware for microscope provided

• Includes instruction manual or equivalent

4.0 Pricing Schedule

Total price for each line must include all required items and incidentals per Section 3.0 of this document.

Line Number Description Quantity Price 0001 Fluorescent microscope & accessories 1 $ 0002 Shipping 1 $

Total contract value: __$__________

5.0 Delivery Location

APHIS PPQ S&T Insect Management and Molecular Diagnostics Laboratory (IMMDL) Moore Air Base, Bldg 6414 22675 N. Moorefield Rd.

Edinburg, TX 78541-5033

6.0 Period of Performance

The estimated period of performance is the Date of Award through 60 days after Award.