As the titles indicates, I would like to search a file in a particular folder, and its subfolders, through Spotlight. Is it possible to do that in Spotlight? If so, how can that be done?
Search in a particular folder and subfolder through Spotlight
searchspotlight
Related Solutions
After some more research, it reminds me that finder's spotlight had boolean UI since OS X 10.5. You can filter for "not a folder" 
for All My Files experience, or with a little bit more filter 
for "all files".
It feels a bit hack-y though.
If you're comfortable opening the terminal you could run the command
locate ABC/readme.txt
and it will give you the full file path to any files on your machine that are named readme.txt in a folder named ABC.
While you're there you can copy and past the path as an argument to the open command and it will open it for you.
Taking it one step further you can use the output of locate as the arguments for open with xargs
locate ABC/readme.txt | xargs open
Will open all files that have ABC/readme.txt on your machine just as if you had double-clicked them.
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Best Answer
Short answer: No, you can not.
Spotlight is designed to be the "central search hub" of your Mac. So whenever you type in something, it will search everything (except "blacklisted" folders and file types that you can set within System Preferences as well as any data from all other user accounts and connected servers unless your account has permission to view them) on your Mac internal storage as well as any additional connected media storage devices.
If you want to search into a specific folder and its subfolders, you will need to go to the actual folder in Finder first. In this example, the selected folder will be "Treehouse"
Then, click on the search bar (typically on the top-right corner of the window) and type in whatever you need to search for. In this example, the term will be "iTunes".
Finally, you will see a gray bar below the Finder toolbar. Select the folder name within the gray bar (if it hasn't been selected already) and it will limit the search to just the files and folders within the said folder.