Question

1. Define and briefly describe in the space below the processes of digital image capture, image enhancement and image analysis in microscopy (1 page limit)

about General and Confocal Microscopy and Image Analysis

Answer 1

GENERAL MICROSCOPY

It is a field at which at which they uses microscopes to view objects and areas of objects that cannot be seen with naked eye.A general microscope consists of mainly an objective lens,ocular lens,lens tube,stage,and a reflector.If an object is placed on the stage which is magnified through the objective lens.When the target is focused,a magnified image can be observed through the ocular lens.This is the principle behind microscopy.

DIGITAL IMAGE CAPTURE

Digital image capturing means digital imaging is increasingly applied to image capture for microscopy.It is an area that demands high resolution,color fidelity and careful management of often limited light conditions.

Why it is more demanding?

The explosive growth in digital imaging is taking the imaging market by storm.The latest digital camers combined with powerful computer software now offer good image quality that is comparable with traditional silver halide film photography.Also they are more easier to use and offer greater flexibility for image manipulation and storage.

PRINCIPLE OR HOW IT IS DONE.

The image is stained with three fluorophores and recorded with a nikon DXM1200 digital camera system coupled to the SMZ1500 steriomicroscope.Then the image was captured by utilizing a combination of fluorescence and Nikon's propietary oblique coherent contrast techniques illumination techniques.A total of four integrations were performed and the image was composed in photoshop to yield the final version of the image.

ADVANTAGES

Images may be simply emailed for consultation and discussion,incorporated into other digital documents,exported to image analysis systems ,it can be posted on a website since they can be easily copied,stored and archived.Can be easily annotated with appropriate software .

IMAGE ENHANCEMENT

By using image enhancement.the visibility of selected features in an image can be improved,but we cannot able to increase the inherent information content.Design of a good image enhancement algorithm should consider the specific features of interest in the microscopic image and the imaging process itself.Image enhancement algorithms are mainly of two types:spatial domain methods and transform domain methods.Spatial domain methods include operations carried out on a whole image or on a local region selected on the basis of image statistics.Main techniques in thus include histogram equalisation,image averaging,sharpening of important features such as edges or contours,and nonlinear filtering.Transform domain enhancement methods manipulate image information in transform domains,such as fourier and wavelet forms.

IMAGE ANALYSIS

Image analysis is the process by the desire to quantify biological charcteristics such as cell count and tissue volume.Image data are acquired and one data set type consists of a series of images corresponding to focal planes looking deeper in the tissue(3D data),and a second type consists of a series of images corresponding to a sequence of the instances imaging a single focal plane.Analysis is done via a combination of segmentation and registration techniques,but is complicated by factors such as live specimen motion during image acquisition.

CONFOCAL MICROSCOPY

Confocal microscopy is a process which creates sharp images of a specimen that would appear otherwise blurred with the conventional microscope and this is achieved by excluding most of the light from the specimen,but not from the microscope's focal plane.

DIGITAL IMAGE CAPTURE

Digitilization of video or electronic image captured through an confocal microscope results in a dramatic increase in the ability to enhance features,extract information,or modify the image.When compared to the traditional mechanism of image capture,photomicography on film,digital imaging and post acquisition processing enables a reversibile,essentially noise - free modification of the image as an ordered matrix of integers rather than a series of analog variations in color and intensity.

Depending upon the illumination conditions ,specimen integrity and preparation ,images captured may require rehabilitation to archive a balance between scientific accuracy ,cosmetic equillibrium and aesthetic composition.Deconvolution is used here,it is a computationally intensive image processing technique that is being increasingly utilized for improving the contrast and resolution of digital images captured in the microscope.

Digital cameras used here are Olympus DP70,Olympus DP-10.

IMAGE ENHANCEMENT

Here for image enhancement deconvolution is used.It refers to the use of algorithms to enhance the signal to noise ratio and resolution of the captured image,typically by reassigning out of focus light back to its orginal position.Specific method applied for deconvolution and the parameters used for the algorithms greatly impact the total image resolution.Mainly it includes both inear and non-linear deconvolutions techniques.

IMAGE ANALYSIS

Analysis is usually by means of scanning techniques whether it is a 2D OR 3D image.The beam is scanned across the sample in the horizontal plane by using one or more oscillating mirrors.The scanning method usually has a low reaction latency and the scan speed can be varied.There are four microscopes for horizontal scanning of the image which include:CONFOCAL LASER SCANING MICROSCOPE,SPINNING DISK,MICROLENS ENHANCED AND PROGRAMMABLE ARRAY MICROSCOPES.