Feasibility of Alugbati (Basella rubra L.

) Fruit Extract as Biological Stain

Abstract

This research project is aimed at producing a natural stain for plant tissues that is inexpensive and whose sources are indigenous and
abundant.

Biological stains make possible the viewing of microscopic plant tissues under microscopes. When stained, the specimens are viewed clearer
and they become more defined. Berries of Basella rubra (alugbati) were crushed using mortar and pestle. The crude extract obtained was
filtered and used as a substitute for crystal violet as primary stain and the extract was used for staining onion and leaf stalk of pechay.

Chapter I: Introduction

Background of the Study:

Biological stains make possible the viewing of microscopic plant and also animal tissues under microscopes. When stained, the specimens
are viewed clearer and they become more defined.
The study of the microscopic details of the structures of plants usually requires some preparation of the material to facilitate observations.
Most of the dyes being used now in biological techniques are synthetic chemical compounds; Natural stains are rarely used. In general,
however, natural or biological stains are superior to synthetic ones in terms of permanence of coloration in microscopic examinations.

Instead of using chemicals for staining such as iodine solution, the researcher though of using a natural means of staining agent such as
malabar spinach or widely known as alugbati, since it shows staining property; it produces a red violet pigment. The researcher aims to
produce a natural stain for plant tissues that is inexpensive and whose sources are indigenous and abundant.

Statement of the Problem:

The study is entitled "The Feasibility of Alugbati Fruit Extract as Biological Stain". Its general objective is to examine the potencial alugbati
extract compared with iodine solution as an affordable and effective staining agent for microscopic observation.

Its specific objective is to find out whether alugbati fruit when used as stain will be effective in clear tracing or observing of onion and leaf
stalk of pechay.

Is the viewing of microscopic plant tissues under microscopes possible?

Hypothesis:
Alugbati fruit extract is a favorable stain for plant cells & tissues. Its effectiveness is eiter comparable or not to the manufactured or
commercial stains.
Significance of the Study:

Laboratories use staining agents to clearly observe specimens. Some schools provide lboratories of their own. But some school laboratories
especially public schools lack some failities and materials needed for concise investigationsand experiments.
In producing this alternative staining agent, it would provide school laboratories with lesser cost conventional stains like iodine solution. The
suggested stain is cheaper than conventional stains and is all naturals that helps in conserving the environment such as reducing water
contamination brought about by improper disposal of used and expired chemical stains. All natural may also mean that it is non-toxic.
Scope and Limitation:
This research project is only limited on the investigation of alugbati fuit extract as biological stain and the said product will only be tested on
plant tissues and cells.
Definition of Terms:

Alugbati- a succulent, branched, smooth, twining her baccous vine, several meters in length. Stems are purplish or green. Leaves are fleshy,
ovate or heartshaped. Fruits are fleshy, stalkless, ovoid or spherical, 5-6cm long and purple when mature.

Stain- a dye or pigment used for staining/coloring cells/tissues' distinctive parts; in determining whether acidic, basic, neutral.

Synthetic- artificial

Chapter II: Review of Related Literature

Alugbati is also known as malabanar spinach, indian spinach with tagalog names, libato and grana, bisayan name, alugbate, bicol name,
arogbati, ilokano names, ilaibakir ang alukbati and has a scientific name f Basella Rubra Linn. Alugbati is aglabrous, succulent, branched vine
over one meter long. The leaves are fleshy, heart-shaped and are demulcent, diuretic and generally used for gonorrhea and balanitis. The
petioes are fleshy, green or purplish. The fruit is ovoid having inside seeds which generally are eaten as dessert. (Plants and Health by
A.C.Sas. 1990)
Alugbati is a vigorous climbing vine in warm climates. This vegetable is very popular in tropical and subtropical areas. Young leaves and tips
are used like spinach in cooking and salad. Seed are sown in spring and early summer. The plant is almost insect-free and it is very easy to
grow. Harvest by cutting the young leaves and tips are vegetable use, but keep 2-3 leaves on the branches for subsequent new branches to
grow.
(http://www.evergreenseeds.com/malabarspinach.html)

Natural stains are best known for its distinct property: permanence of coloration; but due to its arty; most of stains used nowadays are
synthetic. Chemical compounds made from the substance found in coal tar. Yet, natural stains are more superior than them they keep with
specimen for a long time while the synthetic ones easily fade away. Permanence of coloration is important especially for preparations that
require considerable handling over a period of time. Only three natural dyes are still used by biotechnicians: brazilin from brazilwood,
hematoxylin from logwood and hematein from caesal pinacae. (http://www.batobalani.com;vol.22.no.1/)

Chapter III: Methodology

First, materials shoud be gathered by the researcher; 10g of alugbati fruits, mortar and pestle, beaker, sharp razor blade, glass slides,
dropper, glycerin, microscope, 5mL of water, onion, leaf stalk of pechay.

A. Preparation of the Biological Stain

A ten grams of alugbati fruits will be gathered, then, they will be crushed using mortar and pestle in order to produce an average of 250mL
extract. Then, the crude extract will be filtered using a strainer or a filter paper. After that, the researcher placed the filtrate in a clean
container and half of the filtrate will be diluted with water.

B. Testing

Very thin slices of onion and leaf stalk of pechay will be prepared using a sharp razor blade. Sections will be made longitudinally for leaf stalk
of pechay and onion. They will be placed on a glass slides and a drop of alugbati fruit extract with water and without water will be put on
each of the specimens. There will be three replicates in every trial and there will be two trials. After five minutes, a drop of glycerin will be
added. The slides will be examined under the microscope.
C. Observation

D. Analysis of Data

Each of the specimens received four treatments, the first treatment is staining the specimen with alugbati stain without water, second
treatment is staining with alugbati stain with water, third treatment is staining with iodine stain without water and fourth treatment is
staining with iodinestain with water. From the treatments done in two trials, results were obtained. Different treatments were also done to
compare the produced stain with commercial stain to determine their effectiveness.

Chapter IV: Results and Discussion

In this chapter, the researcher will try to show the results and observation of the experiment conducted in a tabulation form.

Table 1.1 Results of the Experiment

SpecimensTreatment 1Treatment 2Treatment 3Treatment 4

Alugbati stain w/ waterAlugbati stain w/o waterIodine stain w/ waterIodine stain w/o water
Leaf Stalk of Pechay 1 3 2 4
Onion 1 3 2 4

The table shows the result of the expriment and compares alugbati stain to the commercial stain/iodine stain. In the set-up, the researcher
had prepared two specimens. Each of the specimens received four treatments. After conducting the experiment, the results are compared to
the control variable which is the commercial stain/iodine stain according to its clarity. The observation were noted and are listed in the table
1.1. After the observation, alugbati stain is feasible and it is comparable to commercial stain in terms of clarity.

Chapter V: Conclusion and Recommendation

Conclusion

In correspond to the collected data and based on the experiments, it was concluded that alugbati stain is feasible and it is comparable to the
commercial stain in terms of clearness but its permanence did not last longer. And also adding water in the stains did not make any
significant difference to its clearness.

Recommendation

The researcher recommends further research on the project especially on the permanence of the stain and its other composition to produce
a better stain.

Acknowledgments

I would like to heartily thank our research teacher Ma'am Balve Granido for the support and supervision and everyone who helped me in this
project.

Bibliography

http://www.evergreenseeds.com/malabarspinach.html

Plants and Health by A.C.Sas. 1990

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