Pollen Calendar, Preparation Method.

1. MADRAS CHRISTIAN COLLEGE, CHENNAI (AUTONOMOUS)
A Brief Presentation On
POLLEN CALENDAR
Submitted By:
Alok Kumar
Reg- 1901711085034

2. POLLEN CALENDAR
 What is a pollen calendar ?
 Types of pollen calendars
A. Pollen Allergy Calendar
B. Crime Scene Pollen Calendar
 Methods of making a pollen calendar
A. Collection of sample
B. Mounting and preparing slide
C. Staining
D. Microscopy
E. Counting Methods
F. Identification
G. Referral Material
H. Constructing Calendar
 References

3. What is a Pollen Calendar?
A pollen calendar is statistical representation which show
pollen peak in atmosphere with respective time period.
A pollen calendar is constructed on basis of field
botanical studies as well as aeropalynological surveys.
The compilation of a pollen calendar is the ultimate
objective of aerobiologists, as this is most useful for
clinicians in diagnosis and treatment of allergy.
“A detailed pollen calendar of a region is a prerequisite
for the immunological treatment of pollen allergies
(Caulton et al., 1997).”
Types of Pollen Calendars
Based on uses , pollen calendar can be defined as various
types ; in which, two types of pollen calendars are main –
 Pollen Allergy Calendar
 Crime Scene Pollen Calendar

4. Pollen Allergy Calendar :
When a pollen calendar is used in field of aero-allergens ,
specially in field of air-borne pollen ; it is known as Pollen
Allergy Calendar .
Crime Scene Pollen Calendar :
When a pollen calendar is used in field of forensic
palynology or solving in crime scene ; it is known as
Crime Scene Pollen Calendar. ( Anna Maria Mercuri ,
Giuliana Trevisan Grandi , Carla Albert Accorsi 2005)
Methods Of Making Pollen Calendar
There are series of processes are involved in preparing a
perfect pollen calendar. These steps are -
A. Collection of Sample
For this collection of pollen sample must be at a
minimum of 3 days per week . To avoid local pollen
sample, in case of aeroallergen , pollen sample collection
from more than one storey height. Such type of sampling

5. can be done by ‘Burkard Volumetric Air Sampler or
Rotorod Rotation impaction Sampler’.
Burkard volumetric sampler is used widely now a days .
This sampler was developed in 1952 by Dr James Hirst. It
draw air 10 liter per hour and any air borne pollen with
sufficient inertia are impacted either on greased tape or
a greased microscopic slide . The impaction surface
moves past the orifice at 2 mm per hour . Wind wane is
there oriented to wind . Standard orifice is sufficient for
particles down to 3.7 micron . It means pollen size more
than this range can be trapped.
Daily pollen counts are reported as daily average pollen
concentrations (pollen grains/m3
) which is the number of
pollen grains divided by the volume of the air sampled
over 24 h.
B. Mounting :
Prepare slides with labeling them with the location, date
and time of collection, also sample number if using more
than one trap. Place a strip of Gelvatol 45 mm long down
the centre of slide . This works as adhesive to hold the

6. tape on slide . The exposed segment of the tape can be
rolled onto Gelvatol using fine forceps and also taking
care not to get bubbles under the tape. Gelvatol should
be allowed to dry for 24 hour before further processing .
Sections can be mounted under 60×24 mm2
.
C. Staining :
Basic Fuchsin can be used directly to stain pollen grains.
Also Calberla’s solution is also being used to stain pollen
grains these days. Acetolysis is always best way for this
pupose.
D . Microscopy
A research grade light microscope with vernier scale is
required for the analysis of pollen . 400x-600x
magnification is enough to identify pollen .
1000x is enough for identifying pollen morphology and
pollen genera can be known.

7. E . Counting Methods :
• Longitudinal Traverse:
Advantages:Provides average prevalence over 24 h and
requires less time to examine a daily slide.
Disadvantages:Due to uneven particle deposition across
the tape, a single traverse may not be representative.
• Twelve Traverses at 4 mm (2 h) Intervals :
The most accurate way of determining the mean pollen
count would clearly be to count all pollen grains on the
24 h trace. However, this would be time consuming and
uneconomic.
F . Identification
Scan each slide using a light microscope to identify
deposited particles and organisms, and record the data
on a specially designed preform a showing the date,
time, location, and type of pollen.

8. Identification of particles should be made at the most
specific classification possible, but pollen can only rarely
be identified to the species level, and some may have to
be placed in groups of multiple genera or families.
Identification is done based on surface morphology of
pollen . Each pollen of each genera have unique
morphology . Therefore this can be said as pollen ID.
G . Referral Material
Consultation of identification manuals are recommended
for descriptions of distinctive features of particular types
of pollen. An atlas may be used to study the geography of
the local area. Literature and monographs may be used
in conjunction with reference slides, to further identify or
confirm the provisional identification.

9. H . Constructing Calendar
Consultation of identification manuals are recommended
for descriptions of distinctive features of particular types
of pollen. An atlas may be used to study the geography of
the local area. Literature and monographs may be used
in conjunction with reference slides, to further identify or
confirm the provisional identification.
Pollen count is measured according to the number of
pollen grains per cubic meter of air. These pollen count
with time period are depicted in form of a chart .
In chart , We present highest density of pollen grain
respective to time period .
We can also depict in chart what kind of allergy caused
by particular pollen grains.
Here are attached some common allergic pollen
dispersing plants -

11. References
 “Pollen Allergy In A Changing World” book
 University Of Worcester studies on allergic
pollen.
 Crime Pollen Calendar –pollen analysis on
corpses throughout one year 2005
 Hyde HA. Atmospheric pollen and spores in
relation to allergy I. Clin Allergy. 1972;2:153–79.
 Kapyla M, Penttinen A. An evaluation of the
microscopic counting methods of the tape in
Hirst-Burkard pollen and spore trap. Grana.
1981;20:131–41.
 Aizenberg V, Reponen T, Grinspun SA, Willeke K.
Performance of Air-O-Cell, Burkard, and Button
samplers for total enumeration of airborne
spores. AIHAJ. 2000;61:855–64

12.  Razmovski V, O’Meara TJ, Taylor DJM, Tovey ER.
A new method for simultaneous
immunodetection and morphologic
identification of individual sources of pollen
allergens. J Allergy Clin Immunol.
2000;105:725–31
 Sterling M, Rogers C, Levetin E. An evaluation of
two methods used for microscopic analysis of
airborne fungal spore concentrations from the
Burkard Spore Trap. Aerobiologia. 1999;15:9–
18.
 Farjon A. World checklist and bibliography of
conifers. Kew: Royal Botanic Gardens; 1998.
 Bottcher MF, Jenmalm MC, Bjorksten B.
Immune responses to birch in young children
during their first 7 years of life. Clin Exp Allergy.
2002;32(12):1690–8.