Accu-Scope 3000-LED series User manual

INSTRUCTIONS

73 Mall Drive, Commack, NY 11725 • 631-864-1000 (P) • 631-543-8900 (F)

www.accu-scope.com • [email protected]

3000-LED

SLIDER PHASE CONTRAST MICROSCOPY

INSTRUCTIONS

PHASE ANNULI

ARE PRE-CENTERED

3000-LED SLIDER PHASE CONTRAST INSTRUCTIONS

Slider Phase Contrast

Installation & Use Instructions

Phase contrast should be used for

observing unstained specimens (i.e.

cultured cells) at high contrast.

Prior to shipment your phase contrast set

was assembled, tested and inspected at

our distribution center in New York. The

phase slider objectives and condenser

were pre-centered and aligned by our

technicians.

IMPORTANT: Before you can use the

phase slider, you must remove the

dust shield cover from the condenser

housing and install the phase annuli

slider in its place. (See FIG. 1)

IMPORTANT: The condenser has

been intentionally installed in a slightly

rotated orientation. This was done to

prevent accidental contact on the

stage control arm when using the

annuli slider.

In the event the condenser is

removed, it MUST BE REINSTALLED

in the original orientation.

The REFERENCE LINE on the

condenser must be ALIGNED WITH

THE SET SCREW in the condenser

holder.

Objectives

Green Filter

Phase Slider

3000-LED SLIDER PHASE CONTRAST INSTRUCTIONS

REMOVING THE

DUST SHIELD COVER

Slide the dust shield cover out from the condenser housing

(FIG. 1). Be sure to keep it to reinstall it should you need to

transport the microscope. FIG. 2 shows the condenser

housing with the dust shield cover removed.

INSTALLING THE

PHASE SLIDER

Before inserting the Phase slider, it is recommended the iris

diaphragm lever be set to PH on the condenser.

Insert the slider into the slot on the condenser as shown in

FIG. 3 & FIG 4.

Move the slider to its position that corresponds to the phase

objective in use (i.e., 40x objective = 40x position on slider).

IMPORTANT!

Raise the condenser to its uppermost position.

Refer to the instruction manual provided with the microscope

on how to focus on a specimen.

FIG. 1

FIG. 2

GREEN FILTER

The included green filter can be mounted on top of the

illuminator housing.

FIG. 3

FIG. 4

Condenser Height Adjustment Knob

3000-LED SLIDER PHASE CONTRAST INSTRUCTIONS

PHASE CONTRAST

MICROSCOPE

The normal microscopic object is seen

because it has regions of varying density. In

normal brightfield illumination a completely

transparent specimen is very difficult to

observe in detail because all areas of the

specimen are equally dense. Darkfield

illumination displays border effects in

completely transparent specimens due to

edge scattering and diffraction of light.

Polarized light is useful when transparent

specimens have directional or crystalline

properties.

Phase contrast microscopy is a type of

illumination system to observe transparent

media. This form of illumination is utilized

extensively in the study of transparent living

cells without the need for staining or fixing

while being able to obtain good image

contrast. The light from phase contrast

illumination arrives at the user’s eyes at ½

the normal wavelength. This light altering

system produces a visible image of an

otherwise invisible, transparent specimen.

The optical light path necessary for phase

contrast is shown in Figure 1. A clear annulus

in the focal plane of the condenser is imaged

at infinity by the condenser and then re-

imaged by the objective in its rear focal

plane. The undiffracted light passes through

this image. It is reduced in intensity and given

a one-quarter wave phase shift by means of

an annular phase pattern in the rear focal

plane of the objective. These two changes in

the undiffracted portion of the beam simulate

the phase and intensity distribution which

would be present in the objective focal plane

if the specimen had density variations rather

than refractive index variations. As a result,

the image formed by the beam interfering

with the diffracted beam simulates that of a

specimen having density variations.

IMAGE FORMATION

BY PHASE CONTRAST

An annular aperture in the diaphragm placed in the focal

plane of the substage condenser controls the illumination

of the specimen. The aperture is imaged by the

condenser and objective at the rear focal plane or at the

exit pupil of the objective. A phase shifting element, or

phase plate, is placed in the image plane. Light passing

through the phase altering pattern acquires a ¼ wave

length advance over that diffracted by the object structure

and passes through that region of the phase plate not

covered by the altering pattern. The resultant interference

effects of the two portions of light form the final image.

Altered phase relations in the illumination rays, induced

by otherwise invisible elements in the specimen, are

translated into brightness differences by the phase

altering plate.

TROUBLESHOOTING GUIDE

PHASE CONTRAST MICROSCOPY

PROBLEM

CAUSE

CORRECTIVE MEASURE

Poor phase

contrast image

is obtained

The condenser phase annulus image

and the objective phase plate are not

aligned

Adjust the phase annulus so that it is

aligned with the objective phase

plate.

The condenser phase annulus and the

objective phase code do not match.

Rotate the phase annulus selector

wheel to the position that matches the

objective in use

The phase difference of the specimen

is too large.

Prepare the specimen using a

different refractive index immersion

fluid

The specimen cover glass is incorrect

Replace with 0.17mm thick cover

glass

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