Education

1. Presentation on “Utilization and
maintenance of Sophisticated analytical
instruments in Research
Dr.Deboja Sharma
Associate Professor
Department of Applied Biology, USTM

2. Research
• Research is a process of systematics inquiry that entails collection of data, documentation of critical information
and analysis and interpretation of that data.
• A research design appropriate for a particular research problem, usually involves the following features.
•
The mean of obtaining information.
•
The objective of the problem to be studied.
•
The nature of the problem to be studied .

3. • Research design have following parts
• Sampling design :Which deals with the methods of selecting items to be observed for the study
•
Observational design :Which relates to the condition under which the observation are to be created.
•
Statistical design:Which concern the question of the of How the information and data gathered are to
be analyzed ?
•
Operational design : Which deals with techniques by which the procedures satisfied in sampling

4. • RESEARCH AREAS IN THE FIELD OF MICROBIOLOGY/BIOTECHNOLOGY

5. Lab
equipment
Laboratory equipment refers to the various tools
used by scientists working in a laboratory.

6. Environmental Microbiology
“Isolation and Characterization of Biosurfactant producing Microorganisms and to determine its invitro
antagonistic activity against Phyto pathogenic fungi.”
Isolation of biosurfactant producing bacteria from oil contaminated soil

7. Incubator
• Is a device used to grow and maintain
microbiological cultures.
• The incubator maintains optimal temperature,
humidity and other conditions such as the
carbon dioxide (CO2) and oxygen content of
the atmosphere inside.

8. Incubator

9. Autoclave
• An autoclave is a pressure chamber used
to sterilize equipment and supplies by
subjecting them to high pressure saturated
steam at 121 °C for around 15–20 minutes
depending on the size of the load and the
contents.
• Used to sterilize culture media, discard, and
other equipments.

10. Autoclav
e

11. HOT AIR Oven
• Device used in sterilization.
• oven uses dry heat to sterilize.
• It used to sterilize items that might be
damaged by moist heat (e.g., glasswares,
powders, oils).

13. Production process of biosurfactant in an orbital shaker incubator
This particular study provides base line data for understanding of the
soil contaminated with crude oil in a particular area. Contamination
of soil environments with crude oil is very hazardous for the human
and other organisms in the ecosystem.

14. TENSIOMETER

15. The basic principle of the Wilhelmy Plate
method
• Wilhelmy plate method refers to a thin plate with known dimensions
that is typically used to measure surface or interfacial tension
between air-liquid or liquid –liquid interface respectively.
• The method is based on measuring the force acting on the plate that
is partially immersed into liquid. The force acting on the plate
F(h)=Pγcos ϴ-ρAhg
P=Perimeter of the plate , γ=Surface tension of the liquid
ϴ=contact angle between the plate and the measured liquid
ρ= density of the liquid, A= surface area of the plate ,g= gravitational
constant

16. • 5 Surface tension measurement
• Surface tension of crude biosurfactant was determined by Wilhelmy plate method in a tensiometer with
respect to distilled water. Average reduction of surface tension was in the pH 4, with the strain X in pH 4
showed the maximum reduction in surface tension as compared to others. Also noted down the variations
in the surface tension of all other strains.
0
10
20
30
40
50
60
70
80
X Y D1 P1 P3
72 72 72 72 72
34
43
34.2
50.8 52
Strains
(pᴴ 4)
0
10
20
30
40
50
60
70
80
X Y D1 P1 P3
72 72 72 72 72
58.6
54.2 53 54.4
61.8
Hours
Surface
Tension(mN/
m)
Strains (pᴴ 6)
0
10
20
30
40
50
60
70
80
X Y D1 P1 P3
72 72 72 72 72
54.4
47.8
53.4
58.4
62.2
Hours
Surface
Tensions(mN/
m)
Strain (pᴴ
8)

18. In all types of microscopes, cell constituents are not
distinguishable, although staining dose , but not
totally.
In fluorescent microscopy, various fluorescent dyes
are used which gives property of fluorescence to only
specific part of the cell and hence it can be focused.

19. Acomponent of interest in the specimen is
specifically labeled with a fluorescentmolecule
called a fluorophore
The specimen is illuminated with light of a specific
wavelength (or wavelengths) which is absorbed by
the fluorophores, causing them to emit longer
wavelengths of light (of a different color than the
absorbed light).
Typical components of a fluorescence microscope
are the light source (xenon arc lamp or mercury-
vapor lamp), the excitation filter, the dichroic
mirror and the emission filter.

20. The illumination light is separated from the
much weaker emitted fluorescence through
the use of an emission filter.
The filters and the dichroic are chosen to match
the spectral excitation and emission
characteristics of the fluorophore used to label
the specimen.
In this manner, a single fluorophore (color) is
imaged at a time. Multi-color images of several
fluorophores must be composedby combining
several single-color images.

25. Fluorescence microscopy is a critical tool for
academic and pharmaceutical research,
pathology, and clinical medicine.

27. Phase contrast microscopy
• It is the first microscopic method
which allow the observation of living
cell.
• It was invented by Frits Zernike and
was awarded noble prize in 1953.

29. Dark Field
Bright Field
Phase contrast
Comparison of Light Microscopy

33. Food & Industrial Microbiology

34. FERMENTER

35. Upstream processing
Upstream processing encompasses any technology
that leads to the synthesis of aproduct.
Upstream includes
= the exploration
= development
= production

36. Downstream
processing
The extraction and purification of a biotechnological
product from fermentation is referred to as
downstream processing.

37. IDEAL FERMENTORPROPERTIES
 Supports maximum growth of the
organism
 Aseptical operation
 Adequate aeration and agitation
 Low power consuming
 Tempurature control system
 pH control system
 Sampling facilities

38.  Minimum evaporation loss
 Minimum use of labour
 Range of processes
 Smooth internal surfaces
 Similar in geometry to both smaller &
larger vessels in pilot plant
 Cheapest material usuage
 Adequate service provisions

39. Provision for control of contaminants
Provision for intermittent addition of
antifoams
Inoculum introduction facility
Mechanism for biomass/ product
removal
Setting for rapid incorporation of
sterile air
Withstands pressure
Ease of manipulation

40. Bioleaching of Alumina from low grade Indian bauxite (41% Al2O3)
by indigenous bacteria with reference to pH, Time and Carbon
source
• The process of metal extraction from low grade ores by using the microorganisims referred as
Bioleaching. It is very simple and effective technique use microorganisms like mesophiles,
thermophilic and extermophiles. The bacteria are mainly Thiobacillusferrooxidans,
Leptospirillum ferrooxidans
• : Mineralogical and Chemical Composition of Bauxite ore containing 41% of alumina.
Chemical ingredients Percentage (%)
Al2O3 41%
SiO2 2.4%
Fe2O3 24.7%
TiO2 2.6%

41. Atomic Absorption Spectrophotometer
•
Atomic Absorption Spectroscopy is a very common technique for detecting metals and
metalloids in samples.
It is very reliable and simple to use.
It can analyze over 62 elements.
It also measures the concentration of metals in the
sample.

42. PRINCIPLE
• The technique uses basically the principle that free
• atoms (gas) generated in an atomizer can absorb radiationatspecific
frequency.
• Atomic-absorptionspectroscopyquantifiestheabsorptionofgroundstateatomsinthe
gaseousstate.
• The atoms absorb ultraviolet or visible light and make
transitionstohigherelectronicenergylevels. Theanalyte
• concentration isdeterminedfrom the amount of
absorption.

44. Schematicdiagramof AAS:

45. ATOMIZERS:
ATOMIZER
FLAME
ATOMIZERS
GRAPHITE TUBE
ATOMIZERS

46. Figure-1: Variation of PH with respect to different Conc. of Carbon Source and Time interval.
Figure-2: Variation of % Al2O3 recovery with respect to time and C- source.
Figure-3: Variation of % Al2O3 recovery with respect to time and C- source.
0
2
4
6
International Research Journal of Environmental Sciences ISSN 2319–1414
Vol. 7(9), 20-27, September (2018) Int. Res. J. Environmental Sci.
12
10
8
1 3 6 15 18 21
PH
9 12
Time Interval in Days
0.3 g ofC Source PH
0.6 g of C SourcePH
0.9 g of C SourcePH
18
16
14
12
10
8
6
4
2
0
1 3 6 15 18 21
%
of
Al
2
O
3
recovery
9 12
Time interval in Days
Al2O3 % in 0.3 C
Source Al2O3 % in
0.6 C Source Al2O3
% in 0.9 C Source
18
16
14
12
10
8
6
4
2
0
1 3 6 18 21
%
of
Al
2
O
3
recovery
9 12 15
Time interval in Days
Al2O3 % in 0.3 C Source
Al2O3 % in 0.6 C Source
Al2O3 % in 0.9 C Source

47. %
of
Al2O3
recovery
0.5
0
1
International Research Journal of Environmental Sciences ISSN 2319–1414
Vol. 7(9), 20-27, September (2018) Int. Res. J. Environmental Sci.
18
16
14
12
10
8
6
4
2
0
7.67 8.35 8.72 9.01 9.22 9.45 9.48
pH
Figure-4: Variation of % of Al2O3 with respect to pH in 0.3g C source.
3
2.5
2
1.5
7.634 8.37 9.01 9.34 9.11 9.36 9.45
%
of
Al2O3
recovery
PH
Figure-5: Variation of % of Al2O3 with respect to pH in 0.6g C source.
Al2O3 %
1
0.5
0
1.5
3
2.5
2
7.631 8.3 8.49 8.74 8.92 9.33 9.64
%
of
Al2O3
recovery
PH
Figure-6: Variation of % of Al2O3 with respect to pH in 0.9g C source.
Al2O3 %

48. Agricultural Microbiology/Biotechnology
• A Study On Phytochemical analysis, Antioxidant activity & Detection of Bioactive compounds
present from the tuber of Stephania glabra.
• The tuber of the plants of Stephania glabra was collected from jungle of Yupia , Arunachal Pradesh on the
September .
• Study of Bioactive Compounds
• 1) Column Chromatography: Column chromatography is the one of the most useful Methods for the
separation and purification of both solids and liquids when carried out small small-scale experiments. The
separation can be liquid /solid (adsorption) or liquid/liquid (partition) in column Chromatography
• PRINCIPLE
• Adsorption
• Mixture of components dissolved in the M.P is introduced in to the column. Components moves depending
upon their relative affinities.

49. COLUMN CHROMATOGRAPHY
•
Adsorption column chromatography, the adsorbent, packed in a glass
column, and a solvent, the mobile phase, that moves slowly through the
packed column. A solvent used as a mobile phase is called an eluent.

50. • A compound attracted more strongly by the mobile phase will move
rapidly through the column, and elute from, or come off, the column
dissolved in the eluent.
• In contrast, a compound more strongly attracted to the stationary
phase will move slowly through the column.

51. • DETECTION OF COMPONENTS
• If the compounds separated in a column chromatography procedure
are colored, the progress of the separation can simply be monitored
visually.
• If the compounds to be isolated from column chromatography are
colorless. In this case, small fractions of the eluent are collected
sequentially in labelled tubes and the composition of each fraction is
analyzed by TLC.

52. Gas Chromatography – Mass spectrometry(GC- MS):
• GC-MS was done out to identify some Bioactive component present in the sample.
• To know the what is bioactive compound are present, 9 highest peak was taken and was measured from the
GC-MS library which was already Present . The common bioactive compounds are PHENOL, 2,4-BIS(1,1-
DIMETHYLETHYL, PHENOL, 3,5-BIS(1,1-DIMETHYLETHYL, 5-EICOSENE, 1-HEXADECANOL, TRIFLUOROACETIC
ACID,N-TRIDECYL ESTER,BEHENIC ALCOHOL, 6H-DIBENZO[A,G]QUINOLIZINE, 5,8,13,13A-TETRAHYDRO-
2,3,9,10-TETRAMETHOXY, BENZENE, 1-METHOXY-4-METHYL-2-(1-METHYLETHYL, BENZENE, 1-METHOXY-4-
METHYL-2-(1-METHYLETHYL), TRIFLUOROACETATE.

53.  Gas chromatography is a technique capable of separating, detecting s
Gas chromatography–mass spectrometry (GC MS) is an analytical
method that combines the features of gas-chromatography and mass
spectrometry to identify different substances within a test sample.
 Gas chromatography is a technique capable of separating, detecting
and partially characterizing the organic compounds particularly
when present in small quantity.
 Mass spectroscopy provides some definite structural information
• from in small quantity.
 e definite structural information
• from in small quantity.

54. Gas
Chromatography
Mass
Spectrometry
Gas Chromatography -
Mass Spectrometry
 =
Identifies (detects) chemicals
based on their molecular weight or mass
A Chemical Analysis Technique
combining two instruments to provide for
powerful separation and identification
capabilities
Separates mixture of chemicals
so each can be identified individually
Gas Chromatography/Mass Spectrometry(GC/MS)

55. 1. Pneumatic controls
2. Injector
3. Oven
4. Column
5. Interface
6. Ion Source
7. MassAnalyser
8. Detector
9. Vacuum System
10. Control Electronics

56. I

57. THANK YOU