
English: 
[Plant Pigment Analysis]
Hello. Thank you for joining me today for
today's video on Plant Pigment Analysis.
My name is Cynthia Sargent, and I'm the biology
curriculum developer here at PASCO scientific.
I want to show you how you can enhance a traditional
lab activity
using two sensors: a Colorimeter and a Spectrometer.
A lot of biology teachers in their classroom
already do a great activity with their students,
which is to separate the pigments from green
spinach leaves
into the different pigments that are actually
in the leaves using paper chromatography.
The paper chromatography allows students to
see that,
yes, there is a green pigment in the spinach
leaves,
but there are also other colors of pigments,
as well.
There's yellow-colored pigments and orange-colored
pigments.
That's a great activity to do with your students.

Chinese: 
[植物色素分析研究]
歡迎您觀賞本段影片
有關於植物色素的分析
我是辛西婭·薩金特，PASCO生物教育專員
我將為您演示如何利用光譜儀與色度計來提升傳統的生物實驗課程
許多生物老師已經發展許多在課堂上應用的實驗活動
例如從綠色菠菜葉萃取色素並利用層析法來觀察色素變化
濾紙層析法讓學生觀察到菠菜葉的色素
綠色、黃色、與橘色等
對學生來說這是個有趣且實用的實驗

English: 
The colorimeter and spectrometer just allow
students to gather additional data
that help make the activity a little more
meaningful for them.
First, I'm going to start with the colorimeter.
I have a colorimeter and a SPARKlink Air that's
attached to my computer here.
I'm going to show you the data in SPARKvue.
I've already calibrated the colorimeter.
It's a simple green-button calibration on
the front of the sensor.
I used a cuvette of ethanol as my blank,
because that's the substance that I extracted
pigments with.
To prepare the pigments for analysis with
the spectrometer and the colorimeter,
I took some leaves, chopped them into little
pieces with some scissors,
and soaked them in ethanol.
You can also use mortar and pestle to grind
the leaves up further.
You can then push the extract through cheesecloth
to get rid of the solid material,
leaving you just with the liquid pigment extract.

Chinese: 
光譜儀與色度計可以幫助學生分析更多數據
有助於學生更進一步探討實驗背後的理論
首先我們將使用比色計來分析
將比色計與SPARKLinkAir連接到電腦
並利用SPARKvue軟體來分析數據
利用比色計上的校準按鈕來校正感應器
在比色皿中倒入乙醇作為標準溶劑
我們將利用乙醇來萃取色素
然後利用光譜儀與比色計來分析
我將一些菠菜葉切碎並浸泡在乙醇溶劑中
您也更進一步利用搗藥杵將其研磨得更細碎
並透過粗萃取來過濾固體物質僅留下液體部分做為樣品

English: 
I've already set up a vial here with my spinach
pigment extract.
Because my colorimeter is calibrated and ready
to go,
I can just take my spinach pigment extract,
place that into the colorimeter,
snap the lid closed, and hit Record.
I've created a page in SPARKvue that shows
the four colors of the colorimeter.
The lights within the colorimeter shine specific
wavelengths of light through that solution.
The choices of light colors are blue, red,
green, and orange.
For the plant pigment analysis,
I like for students to look at all four of the colors for comparison.
You could also have them look at transmittance.
You can set SPARKvue up to measure a specific
color,
and you can measure the absorbance or transmittance
of that color.
In an analysis of plant pigments, we're going
to relate the data to photosynthesis.
So what we're really interested in is what colors
of light are the leaves absorbing?

Chinese: 
我已經製備好一小罐菠菜萃取樣品
色度計已經校正完畢並準備就緒
所以我可以將菠菜萃取樣品放到比色計中
蓋上蓋子並開始記錄
利用SPARKvue創建一個頁面可以同時顯示比色計四種顏色
比色計可以設定四種不同的光源的燈光通過溶液
分別是：藍光、紅光、綠光、以及橙光
對於植物色素的分析我比較喜歡讓學生直接觀察四種顏色的變化
您也可以讓學生看看透光率
您可以在SPARKvue軟體中，針對某種色光測量穿透率與吸收率
在植物色素的分析中我們要的數據涉及的光合作用

Chinese: 
所以我們對植物葉子會吸收什麼顏色的光非常感興趣
這意味著植物對該色光的利用情形
如果光被反射則表示該色光無法被植物所利用
四種顏色當中藍光與紅光的吸收率較高
而綠光與橙光的吸收率較低
這有助於學生了解在層析紙上所見的顏色為什麼是綠色與黃色
實際上是被反射的色光
利用率較低的色光
而植物對於藍光與紅光的利用率相對較高
並獲得光合作用的能量
比色計所測量到的大數據可以有效幫助學生透過色光的分析來理解色素的組成
光譜儀則提供波長範圍更廣的分析
那麼該如何利用光譜儀呢

English: 
If they're absorbing that color of light,
then that means that the plants are actually
going to make use of that color of light.
If it's being reflected back, then the plants
aren't able to use that color of light.
Of the four colors, blue and red show the
highest absorbance.
Green and orange show the lowest absorbance.
This would help students understand that the
green pigments that make the leaf green,
or the yellow pigments that they see on the
chromatography paper,
are actually reflecting those colors.
They're not absorbing much of that.
It's the blue and the red that is going to
be the most useful to the plant
to obtain energy for photosynthesis.
With the colorimeter you get great data
that can enhance the chromatography setup
that you normally have students go through.
The spectrometer will provide a broader range
of wavelengths.
What you're able to do with the spectrometer

English: 
is get the complete absorbance spectrum for
the pigments in leaves,
rather than just a snapshot look at those
four different colors that the colorimeter provides.
For the spectrometer, I'm going to show you
the spinach extract again,
and we're also going to compare it to plums.
I used the same method of cutting the leaves
into tiny pieces and soaking them in some ethanol
to be able to get an extract of the pigments
present in these purple-colored plum leaves.
I'm going to switch to PASCO's Spectrometry
app.
There are options listed at the top to do
a number of different things.
In this case, we're interested in analyzing
a solution.
We've got our pigments dissolved in ethanol,
and we want to see the absorbance of all of
these visible light wavelengths.
Before I get started, I need to calibrate
the spectrometer.
There are two calibration buttons near the
Record button.

Chinese: 
你可以得到一個完整的植物色素吸收連續頻譜
不僅僅只針對色度計所提供四個固定的光源
針對光譜儀的分析我們將再次利用菠菜萃取樣品
與李子萃取樣品做比對
以同樣的方法將李子萃取物浸泡於乙醇之中
能夠得到紅紫色的色素萃取樣品
接著我們打開PASCO光譜分析軟體
在軟體主畫面上部有許多不同的分析模式
我們選取溶液分析模式
我們有溶解在乙醇的萃取樣品
我們希望能看到可見光吸收波長的頻譜分布
開始之前我們需要先校準光譜儀
在畫面下部有兩個校準按鈕

English: 
First I calibrate dark.
So I take my reference solution --
again, it's just ethanol, since that's what
I extracted my pigments in --
and I press Calibrate Dark.
Then I press Calibrate Light.
The spectrometer is turning the internal lights
off during the calibrate dark procedure,
and then it's turning the lights on in the
calibrate light procedure.
Now that my spectrometer is calibrated,
I'm going to take my spinach pigment extract,
put that into the spectrometer, and hit Record.
You can see a higher absorbance of the blue
light and the red light.
I can get a little better view of this data
by clicking and dragging on the axes here.
Biology teachers definitely wouldn't be too
surprised at this result --
again, the peaks being in the blue and red
wavelength ranges --
but this is really cool for students,

Chinese: 
首先校正暗光源
取出裝有乙醇的比色皿當作標準溶液
點擊暗光源校準鍵
然後點擊亮光源校準鍵
暗光源校正過程中光譜儀會先關閉光源
然後在亮光源校正過程中會自動開啟光源
現在光譜儀已經完成校正程序
取出菠菜萃取色素將其置入光譜儀當中
並且點擊紀錄鍵
我們可以觀察到在藍光與紅光區域有較高的吸收率
拖曳座標軸可以調整尺規以得到更清楚的顯示範圍
生物老師對這個結果肯定不是太驚訝
在藍光與紅光區域有較高的吸收率
但是對學生來說這卻是非常有趣

English: 
because typically they only get this view
of data from a picture in their textbooks.
For them to extract pigments from real leaves
and put it into a spectrometer
and get the absorbance spectrum themselves
is a really meaningful activity for them.
It's easy to compare different leaves.
It's really easy to send students into inquiry,
following the spinach procedure.
They can bring in all different kinds of leaves,
extract the pigments like those in the plum,
and easily compare different kinds of leaves
to each other.
I put the plum pigment extract into the spectrometer.
I'm going to hit Record again.
We'll see where that peak is there, in the
blue.
You can see still peaks at the blue and the
red,
but if I stop data collection and turn on
the Comparison tool,
I can see both absorbance spectra at the same
time.
You can see that the solution two --
which I'm going to rename Plum --

Chinese: 
因為他們通常只能從書本上閱讀與光合作用相關的理論
所以對學生來說自己動手萃取植物色素並分析吸收光譜
這非常有意義且可以幫助理解
透過光譜儀可以很容易的分析不同植物葉子萃取物之間色素的差異
例如菠菜萃取物與李子萃取物的顏色色素與吸收光譜的比較
將李子萃取樣品置入光譜儀中並點擊紀錄
我們可以觀察到最大的吸收峰值落在藍色的區域
而在紅色區域有另一個叫小的吸收峰值
若我停止計錄並打開比較工具
我可以同時看到兩個吸收光譜
可以看到溶液#2
我們可以重新命名為"李子萃取物"

Chinese: 
這兩種萃取物的光譜有明顯的差異性
菠菜在藍光與紅光區域有較高的吸收率
而在綠光與黃光區域的吸收非常低
而李子萃取樣品吸收更多的綠光與黃光
所以其葉子呈現較深的顏色而非綠色
而且也反射更多的紅光而呈現紅色
而在藍光區域的吸收率是相當大的
所以光譜儀可以幫助學生實際觀察到萃取色素的吸收光譜
也可以用來分析不同的光源進一步應用在光合作用之上

English: 
has a couple key differences when compared
to the spinach.
The spinach, again, had those peaks at blue
and red,
but very low absorbance at green and yellow
wavelengths.
The plum absorbs a lot more of the green and
the yellow.
It's reflecting less of those colors, making
it look less green than the spinach.
It also has a lower absorbance of red,
so it's also reflecting more red light than
the spinach.
It has a higher absorbance in the blue light
wavelength.
The spectrometer, again, is great to help
your students see for themselves
the absorbance spectrum for a collection of
pigments extracted from leaves,
but you can also extend the inquiry into photosynthesis
by using the spectrometer to analyze different
light sources.
In the next part of the video, we're going
to take a look

English: 
at how you can obtain the emission spectra
for different light sources.
[Analyzing Light Sources]
In the next part of this video, we're going
to look
at how we can use the spectrometer to analyze
different light sources.
This can be another great extension to your
study of photosynthesis with your students.
I've replaced the use of cuvettes in the spectrometer
with a fiber optic cable.
Instead of analyzing a solution like we did
for the pigment extract,
now we want to analyze a light source.
In the PASCO Spectrometry app, I've selected
Analyze Light.
It takes me to a new page within the application.
Now, on my y-axis, it shows intensity.
When I click the Record button
and hold the fiber optic cable just directly
underneath that light source,
I get a good view of the wavelengths that
are being emitted from that light source.

Chinese: 
在下一段的影片當中我們將利用光譜儀分析不同光源的發射頻譜
[光源頻譜分析]
在這一階段的影片中我們將探討如何利用光譜儀來分析不同的光源
這可以幫助學生進行另一個關於光譜更進階的研究
我將光纖連接到光譜儀的樣品槽
有別於植物色素分析利用溶液分析模式
現在我們將啟動光源分析模式
在PASCO光譜分析軟體介面中選取光源分析模式
軟體會新建一個分析頁面
而縱軸會顯示光照的強度
當我點擊紀錄鍵並將光纖對準待測光源
可以得到一個光源的波長分布圖

Chinese: 
點擊停止健可以記錄下光源的分布頻譜
然後利用光纖對準不同光源重複這個過程
接著開啓對比功能可以將兩次紀錄同時顯示在一張圖上
調整尺規可以得到較清楚的視圖
可以發現這兩個光源的頻譜非常相似
主要是藍光、綠光、以及橙光
同時可以看到一些重要的區別
如果我們放大藍光區域
可以發現第二個光源在藍光區域的強度大於第一個燈泡
如果觀察黃光區域

English: 
I can stop recording so I get a record of
that light source's emission spectrum.
Then I can move the fiber-optic cable to a
different light source and repeat the process.
Then, using Comparison mode, I can look at
those two together
and scale my axes a little bit here to get
the greatest view of my data.
You can see that the main wavelengths
emitted are similar for these two light sources.
We get some blue light, mainly green light,
and orange light.
You can also see a few important differences.
If we look a little bit closer at the blue
light part of the spectrum, the second light bulb,
source two here, had a greater amount of light
emitted in the blue wavelengths.
If we move over and analyze the yellow part
of the spectrum,

Chinese: 
則第二個光源較第一個光源的光強低上許多
而在橙光區域兩個光源的強度有極大的差異
第一光源散發出大量的橙色的光
而第二個光源強度較弱
兩個光源都幾乎是白熾光
外觀來看其中一個光源略顯暖白，另一個則略顯冷白
這可以讓學生預測兩種光源差異對於光合作用速率的影響
您可以讓學生利用兩種方式來進行實驗來研究光合作用速率
將菠菜葉置入代謝瓶中並利用二氧化碳感應器來測量植物的吸收速率
請參考我們的"光合作用和呼吸作用"影片

English: 
that second light source was emitting less
yellow light than the first light source was.
Then we can see a big difference in the orange
part of the spectrum.
You can see that first light source is emitting
a lot of orange light,
and the second light source is emitting less
of that.
Comparing two white light sources with your
students, they both look white,
although students might see that this one's
a little more yellow,
this one's a little more blue in just the
appearance to their eyes.
This can be the starting point for them to
make predictions
about what the rate of photosynthesis would
be for each of these two light sources.
You can have your students go perform experiments,
using one of two main methods for getting
photosynthetic rate.
One would be to place spinach leaves in a
bottle
and use a Carbon Dioxide Sensor to measure
the rate
at which those leaves take in carbon dioxide.
You can see a procedure like that in our video
titled "Photosynthesis and Respiration".

English: 
You can also use an experiment that is commonly
done in advanced biology classes,
using the chemical DPIP -- a blue-colored
chemical --
to measure photosynthetic rate.
The DPIP will accept excited electrons that
are produced when light is absorbed by pigments
and will turn colorless when it accepts those
electrons.
If you place these cuvettes with DPIP and
chloroplasts under a light,
then the color will decrease over time.
The colorimeter is a great way to quantify
that change in color over time.
This is a great way to get your students thinking
about light sources and plant pigments
and to relate the data from different sources
--
a colorimeter, a spectrometer, a carbon dioxide
sensor --
and be able to bring the data from those different
sources together
for good understanding of photosynthesis and
what affects that photosynthetic rate.
Thank you for joining me today.

Chinese: 
您也可以利用生物學相關實驗套組
利用化學DPIP來測量光合作用速率
DPIP會吸收光被色素吸收所產生的激發電子而轉變為無色
如果您將葉綠體樣品與DPIP同置於光源之下
DPIP的顏色會隨時間變淡
而色度計是量化的顏色隨著時間的推移變化的好辦法
這是一個偉大的方式讓學生思考植物色素與光合作用的關聯
並同時測量各種不同數據
諸如：色度計、光譜儀、二氧化碳感應器等
最後將這些數據整合在一起
幫助學生深刻理解光合作用與其速率的影響變因
謝謝觀賞

Chinese: 
有任何問題請洽台灣地區總代理 
錫昌科技 0800-026677
