Praktikum Mikroskop

Peni KS Mutalib

Goals

• Being skilled to use microscope

• To know some important principle so that student could operate microscope: good in short time.

• Determine alternatif procedure which be wanted

• Making adaptor self and taking photo (optional)

Assignment in Practice Microscope January 2008

1. Mention the parts of a microscope!2. What is the function of those parts?3. How to optimalize the functions?4. What should be done to maximize your RP?5. What is the NA of your microscope?6. And from your right and left hand side microscope? 7. RP of no. 5 and 6: what is better? What is your argument?8. Figure the tissue with low, medium and high power

(magnification of objective 10x, 40x, and 100x)! Write the overall magnification 100x, 400x, 1000x.

9. Report your experiences in using microscope: The steps!10.What is the difference with the steps mention in theory?

Micoscope Figure

• Oculer: 10x, 15x, 20x• Objektive 10x, 40 x, 100x dan NA 0,45,

0,65• Condensor • Light source or mirror• Immersion oil• Magnification• Object glass

Optical Pathways (Magnifying)

Several theory

• Resolving Power

• NA: Numerical Aperture

• Immersion oil

• How to use Condensor

Lens protection and putting slide while

• Bringing microscope

• Cleaning lens

• Focusing small objective power 4x and medium 10x

• Studying slides with high power 40-45x or when using immersion oil

• Rapikan mik setelah selesai !

Identify part of the microscope• Do it several times, think about the

function

Different location of coarse and fine adjustment

Fine ajustment

How to get the best RP from a microscope incl. how and when to

do:

• Cleaning the lenses

• Illuminate the specimen

• Focusing the condenser

• Ajust the diagfragma

• Using immersion

• Setting light intensity

Ocular and objective lens

Magnification & Numeric Aperture

NA = n sin

Resolving Power

Resolution of lens camera is revealed by how many lines per inch (lpi)

Lens resolution• Is the ability of a lens to free shadow from

2 point of thing that is very close.• Resolusion limit = 1.22/D (Resolusion depend

on )

• Daya urai (=Resolving Power;RP;s)= f= 1.22f/D (D=2sin) where f=jarak objek ke lensa objektif

How smaller how smaller RP

How larger D how smallerRP

How bigger how smaller RP

Light source

Long wave length have greater energy

Electromagnetic wave spectrum

Step in focusing microscope

Step

Step

The best RP visual microscope +- 200 nm

Magnification is not the same with RP

RP microscope mainly depend on obj lens (oculer-max 10x, only magnify image which is found from obj lens-max 100x). Oculer lens didn’t make better the resolusion. Higher magnification with low resolution image blur which is < function

NA= n sin

RP=K.

NA

L objOn the objective was wrote Plan 25/0.45 (Magnification/NA) 160/0.17Tube length in mm

Thickness(in mm) deck glass for which obj lens is corected (dry field)

RP = 0,61 NA

=(0,61)(5,50 x 10-7 m

1,62 x 10 –7 m 200

nmn=1,8 (n immersion oil)

sin 0,90

Large (Fast) NA vs. Small (slow) NA

Thus, NA:

Condensor

• Make better RP with making light lamp 100% of the image-forming kind

• How: with focusing light in the shape of inverse cone, with apex at the specimen and base at the objective

Modern Research Microscope

Tempo, 22 Juni 2003: hal. 16

NA 1.40

CCD camera US$3500

MPixel ?