Formal Report

Exercise 1
The Microscope: Principles, Parts, Use and Care

I. Introduction

The human eye can is an optical system having a resolving of 0.1 mm. If two objects are
separated by a gap of less than 0.1 mm, they would appear as one. Objects with dimension less than
0.1 mm cannot be anymore seen by the human eye and will become microscopic. A microscope can
be simple or compound. The simple microscope, invented by Anton van Leeuwenhoek, has only
one lens. Simple microscope is no longer used today. The compound microscope, invented by
Zacharias Janssen in 1595, has more than one lens.

Light compound microscope is the most familiar laboratory instrument in the field of
microbiology. Its optical system consists of aligned lenses work together to magnify or enlarge the
image of the specimen under the microscope. The objective lens magnifies the image of the
specimen. The image produced is magnified further by a second lens called the ocular, resulting to a
much enlarged virtual image. The visible light of the electromagnetic spectrum, ranging from 400
nm to 700 nm is used to illuminate this microscope. The light compound microscope also has its
limitations. Viruses with dimensions lower than the resolving power of light compound microscope
can be viewed under an electron microscope.

This exercise aims to compute the microscopic parameters like total magnification and
resolving power of each objective using Abbes equation and to know their practical application.
The resolving power or limit resolution of a microscope corresponds to its ability to refine the
details of a specimen. The resolving power is more important in determining the usefulness of a
microscope than its magnification.

II. Methodology

The limit of resolution was computed using Abbes Ernst Abbe equation:

where (lambda) is the wavelength of light used by the microscope; (eta) is the the
refractive index of the medium between the objective and the glass slide and (theta) is
half of the value of angular aperture, also known as cone angle.

The barrels of each objectives were observed. The numbers written on each barrel represent
its magnification, numerical aperture, tube length and the ideal thickness of the cover slip to be
used. The values of NA or numerical aperture and refractive index where noted from each barrels.
The standard refractive index of air is 1.00, which is applicable for LPO and HPO. The refractive
index of oil in OIO is 1.50.

III. Results and Discussion

Table 1.
Microscopic Parameters
LPO
 HPO
OIO
Numerical Aperture (NA)
0.25
0.65
1.25
Half of Angular Aperture
14.48
40.54
56.44
Angular Aperture
28.96
81.08
112.89
Resolving power in um
1.34
0.52
0.27

An object cannot be observed under the objective lens when its dimensions are below the
specific limit of resolution. In this case, a 0.08mm object can be viewed under all objectives
because its converted value, 80m, is higher than 1.34m, 0.52m and 0.27mthe
resolving powers of LPO, HPO, and OIO respectively. The same case is also applied to a
0.01mm object, which is then converted into 10m. Meanwhile, a 0.29m object cannot be
viewed under LPO and HPO because 0.29m is lower than the values of their limit of
resolutions, 1.34m and 0.52m. The same case is also applied to a 0.15m object; it
cannot be seen under the objectives because 0.15m is lower than 1.34m, 0.52m and
0.27mthe resolving powers of LPO, HPO, and OIO respectively.

Table 2.
Can you see the following microscopic objects under each objective?
Dimension of Observed Object
LPO
HPO
OIO
0.08 mm



0.01 mm



0.29 m



0.15 m

An object cannot be observed under the objective lens when its dimensions are below the
specific limit of resolution. In this case, a 0.08mm object can be viewed under all objectives
because its converted value, 80m, is higher than 1.34m, 0.52m and 0.27mthe
resolving powers of LPO, HPO, and OIO respectively. The same case is also applied to a
0.01mm object, which is then converted into 10m. Meanwhile, a 0.29m object cannot be
viewed under LPO and HPO because 0.29m is lower than the values of their limit of
resolutions, 1.34m and 0.52m. The same case is also applied to a 0.15m object; it
cannot be seen under the objectives because 0.15m is lower than 1.34m, 0.52m and
0.27mthe resolving powers of LPO, HPO, and OIO respectively.

IV. Conclusion

Abbes equation is essential in computing for the resolving power of each objective. Any
value less than the calculated resolving power will not be seen under the said objective. It can be
resolved by using another objective, therefore increasing the value of NA or by lowering the value
of lambda. Filters may also be used to see the specimen being observed.

V. References